Substituted (1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9h-purin-8-yl) phenyl derivatives, their preparation and their use in the treatment of inflammatory conditions and immune disorders

ABSTRACT

The present invention provides a compound of formula (I)                    
     or a solvate thereof wherein: 
     X is —O— or —NH—; 
     Q is (—CH 2 —) p , (—CH═CH—) p , (—C≡C—) p  where p is an integer of from 0 to 4; 
     R 1  is hydrogen or methyl; 
     R 2  and R 3  independently represent O or S 
     n is an integer of 1 to 50; and 
     R is hydrogen or methyl. 
     The present invention also provides pharmaceutical compositions and methods of treatment using the compounds of formula (I).

RELATED APPLICATIONS

This Application is a continuation of U.S. patent application Ser. No.09/367,313, now U.S. Pat. No. 6,355,646, filed Dec. 22, 1999 which wasfiled pursuant to 35 U.S.C. §371 as a United States National PhaseApplication of International Application No. PCT/EP98/00784, filed Feb.12, 1998 and published as WO 98/35966, Aug. 20, 1998, which claimspriority to Great Britain Application No. 9703044.9, filed Feb. 14,1997.

The present invention relates to complex esters and amides of phenylxanthine derivatives, processes for their preparation, pharmaceuticalformulations comprising them, and their use in medicine, particularly inthe prophylaxis and treatment of septic shock, inflammatory conditions,as well as immune disorders.

Septic shock is induced by means of a complex series of events involvingmany different pathways and mediators of disease response (see forinstance, The Lancet, Vol. 338 (1991), p732-739, and Annals Of InternalMedicine Vol. 115 (1991), p457-469), including, inter alia, products ofarachidonic acid metabolism and platelet aggregation.

The adhesion of circulating leukocytes to the vascular endothelium is acrucial event in the pathogenesis of inflammatory responses.Inflammatory and immune mediators can stimulate the adhesion process byincreasing the adhesiveness of the leukocyte or the endothelial cellthrough the activation, upregulation, or induction of various adhesionmolecules on the cell surface.

Anti-inflammatory drugs currently available have limited efficacy, oftenwith side effects. Monoclonal antibodies used experimentally foranti-adhesion therapies have theoretical disadvantages for treatment ofchronic diseases. Therefore, the discovery and development of smallmolecules which specifically block or inhibit the adhesive interactionsof leukocytes and the endothelium is an attractive area of therapeuticintervention.

PCT Application No. GB 9501808 describes compounds of formula:

Wherein

m and n are independently integers from 0 to 10;

X and Y are independently oxygen or sulphur;

(—Q—) is (—CH₂—)_(p) or (—CH═CH—)_(p) where p is an integer of from 1 to4; and

A and B are independently methyl, branched C₃₋₆ alkyl, C₃₋₈ cycloalkylor C₃₋₈ cycloalkenyl;

and salts, solvates and pharmaceutically acceptable esters and amidesthereof; and their use in treatment of septic shock, allergic, andinflammatory conditions. The compounds had been found to inhibit one ormore of the enzymes 5-lipoxygenase, cyclooxygenase, and lyso-PAF:acetyl-CoA acetyltransferase.

We have now surprisingly discovered a series of complex esters andamides of selected phenyl xanthine derivatives which inhibit theexpression of adhesion molecules on human umbilical vein endothelialcell (HUVEC) monolayers at very low concentrations and which aretherefore indicated for treatment of inflammatory conditions and immunedisorders.

DETAILED DESCRIPTION OF THE INVENTION

Accordingly the present invention provides a compound of formula (I):

or a solvate thereof wherein:

X is —O— or —NH—;

Q is (—CH₂—)_(p), (—CH═CH—)_(p), (—C≡C—)_(p) where p is an integer offrom 0 to 4;

R¹ is hydrogen or methyl;

R² and R³ independently represent O or S.

n is an integer of 1 to 50; and

R is hydrogen or methyl.

According to a further aspect, the present invention provides a compoundof formula (I) as defined above wherein X is —O— or —NH— and R¹ is H; ofthese, compounds wherein n is an integer of 8 to 20 are preferred, andthose wherein n is an integer of 8 to 15 are more preferred.

Conveniently R³ represents O and R² represents O or S but morepreferably R³ and R² both represent O.

According to a further aspect of the invention, P preferably represents0 or 1.

According to a further aspect, the present invention provides a compoundof formula (I) as defined above wherein Q is (—CH═CH—)_(p).

Preferably the

substituent is attached to the phenyl ring in the para position.

The invention also includes mixtures of compounds of formula (I) in anyratio, for example wherein n varies within the same sample.

A particular subgroup of compounds is of formula 1a

or a solvate thereof wherein:

X is —O— or —NH—;

Q is (—CH₂—)_(p) or (—CH═CH—)_(p) where

P is an interger from 1 to 4;

R¹ is hydrogen or methyl

n is an integer of 1 to 50; and

R is hydrogen or methyl.

Particularly preferred compounds of the invention include

(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Decaethylene Glycol Methyl Ether Ester;

(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Nonaethylene Glycol Methyl Ether Ester;

(E)-3-[1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]cinnamicAcid Nonaethylene Glycol Methyl Ether Ester;

(E)-4-[1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicacid Nonaethylene Glycol Methyl Ether Amide and

(E)-4-[1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]benzoicacid Nonaethylene Glycol Methyl Ether Ester

or a solvate thereof.

The compounds of the present invention are capable of existing asgeometric and optical isomers. All such isomers, individually and asmixtures, are included within the scope of the present invention. WhereQ contains a double bond, compounds in the form of the E-geometricisomers are preferred.

As mentioned hereinbefore, compounds of formula (I) and solvatesthereof, have use in the prophylaxis and treatment of inflammatoryconditions and immune disorders, as demonstrated hereinafter in thebiological assays in which representative compounds of the presentinvention have been shown to be active.

Examples of inflammatory conditions or immune disorders are those of thelungs, joints, eyes, bowel, skin, and heart; particularly thoseassociated with the infiltration of leucocytes into inflamed tissue.Conditions of the lung include asthma, adult respiratory distresssyndrome, bronchitis and cystic fibrosis (which may additionally oralternatively involve the bowel or other tissue(s)). Conditions of thejoint include rheumatoid arthritis, rheumatoid spondylitis,osteoarthritis, gouty arthritis and other arthritic conditions.Inflammatory eye conditions include uveitis (including iritis) andconjunctivitis. Inflammatory bowel conditions include Crohn's disease,ulcerative colitis and distal proctitis. Skin diseases include thoseassociated with cell proliferation, such as psoriasis, eczema anddermatitis (whether or not of allergic origin). Conditions of the heartinclude coronary infarct damage. Other inflammatory conditions andimmune disorders include tissue necrosis in chronic inflammation,endotoxin shock, smooth muscle proliferation disorders (for example,restenosis following angioplasty), and tissue rejection followingtransplant surgery.

Accordingly, the present invention provides a method for the prophylaxisor treatment of an inflammatory condition or immune disorder in amammal, such as a human, which comprises administration of atherapeutically effective amount of a compound of formula (I), or apharmaceutically-acceptable solvate thereof. The present inventionfurther provides a method for the prophylaxis or treatment of septicshock in a mammal, such as a human, which comprises administration of atherapeutically effective amount of a compound of formula (I), or apharmaceutically acceptable solvate thereof

In the alternative, there is also provided a compound of formula (I), ora pharmaceutically acceptable solvate thereof for use in medicaltherapy; particularly, for use in the prophylaxis or treatment of aninflammatory condition or immune disorder in a mammal, such as a human.The present invention further provides a compound of formula (I), or apharmaceutically acceptable solvate thereof for use in the prophylaxisor treatment of septic shock.

The amount of a compound of formula (I) or pharmaceutically acceptablesolvate thereof, which is required to achieve the desired biologicaleffect will depend on a number of factors such as the use for which itis intended, the means of administration, and the recipient. A typicaldaily dose for the treatment of septic shock, for instance, may beexpected to lie in the range of 0.005 mg/kg-100 mg/kg, preferably0.5-100 mg/kg, and most preferably 0.5-20 mg/kg. This dose may beadministered as a single unit dose or as several separate unit doses oras a continuous infusion. An intravenous dose may be expected to lie inthe range of 0.0025 mg/kg to 200 mg/kg and would typically beadministered as an infusion. Similar dosages would be applicable for thetreatment of other disease states. For administration to the lungs of asubject by aerosol an amount of the compound should be used sufficientto achieve concentrations on the airway surface liquid of the subject ofabout 2 to 1000 μmol.

Thus, in a further aspect of the present invention, there are providedpharmaceutical compositions comprising, as active ingredient, a compoundof formula (I) or a pharmaceutically acceptable salt or solvate thereof,together with at least one pharmaceutical carrier or recipient. Thesepharmaceutical compositions may be used in the prophylaxis and treatmentof conditions such as septic shock, inflammatory conditions, and immunedisorders. The carrier must be pharmaceutically acceptable to therecipient and must be compatible with, i.e. not have a deleteriouseffect upon, the other ingredients in the composition. The carrier maybe a solid or liquid and is preferably formulated as a unit doseformulation, for example, a tablet which may contain from 0.05 to 95% byweight of the active ingredients. If desired other physiologicallyactive ingredients-may also be incorporated in the pharmaceuticalcompositions of the invention.

Possible formulations include those suitable for oral, sublingual,buccal, parenteral (for example subcutaneous, intramuscular, orintravenous), rectal, topical including transdermal, intranasal andinhalation administration. Most suitable means of administration for aparticular patient will depend on the nature and severity of thecondition being treated and on the nature of the active compound, butwhere possible, iv administration is preferred for the treatment ofseptic shock, for instance. For the treatment of a condition such asasthma, however, oral or inhalation, would be the preferred route ofadministration.

Formulations suitable for oral administration may be provided asdiscrete units, such as tablets, capsules, cachets, lozenges, eachcontaining a predetermined amount of the active compound; as powders orgranules; as solutions or suspensions in aqueous or non-aqueous liquids;or as oil-in-water or water-in-oil emulsions.

Formulations suitable for sublingual or buccal administration includelozenges comprising the active compound and, typically a flavoured base,such as sugar and acacia or tragacanth and pastilles comprising theactive compound in an innert base, such as gelatine and glycerine orsucrose acacia.

Formulations suitable for parenteral administration typically comprisesterile aqueous solutions containing a predetermined concentration ofthe active compound; the solution is preferably isotonic with the bloodof the intended recipient. Although such solutions are preferablyadministered intravenously, they may also be administered bysubcutaneous or intramuscular injection.

Formulations suitable for rectal administration are preferably providedas unit-dose suppositories comprising the active ingredient in one ormore solid carriers forming the suppository base, for example, cocoabutter.

Formulations suitable for topical or intranasal application includeointments, creams, lotions, pastes, gels, sprays, aerosols and oils.Suitable carriers for such formulations include petroleum jelly,lanolin, polyethyleneglycols, alcohols, and combinations thereof Theactive ingredient is typically present in such formulations at aconcentration of from 0.1 to 15% w/w.

Formulations of the invention may be prepared by any suitable method,typically by uniformly and intimately admixing the active compound withliquids or finely divided solid carriers or both, in the requiredproportions and then, if necessary, shaping the resulting mixture intothe desired shape.

For example a tablet may be prepared by compressing an intimate mixturecomprising a powder or granules of the active ingredient and one or moreoptional ingredients, such as a binder, lubricant, inert diluent, orsurface active dispersing agent, or by moulding an intimate mixture ofpowdered active ingredient and inert liquid diluent.

Aqueous solutions are typically prepared by dissolving the activeingredient in saline to which cyclodextrin has been added.

Suitable formulations for administration by inhalation include fineparticle dusts or mists which may be generated by means of various typesof metered dose pressurised aerosols, nebulisers, or insufflators.

For pulmonary administration via the mouth, the particle size of thepowder or droplets is typically in the range 0.5-10 μm, preferably 1-5μm, to ensure delivery into the bronchial tree. For nasaladministration, a particle size in the range 10-500 μm is preferred toensure retention in the nasal cavity.

Metered dose inhalers are pressurised aerosol dispensers, typicallycontaining a suspension or solution formulation of the active ingredientin a liquefied propellant. During use, these devices discharge theformulation through a valve adapted to deliver a metered volume,typically from 10 to 150 μl, to produce a fine particle spray containingthe active ingredient. Suitable propellants include certainchlorofluorocarbon compounds, for example, dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane and mixtures thereof.The formulation may additionally contain one or more co-solvents, forexample, ethanol surfactants, such as oleic acid or sorbitan trioleate,anti-oxidants and suitable flavouring agents.

Nebulisers are commercially available devices that transform solutionsor suspensions of the active ingredient into a therapeutic aerosol misteither by means of acceleration of a compressed gas typically air oroxygen, through a narrow venturi orifice, or by means of ultrasonicagitation. Suitable formulations for use in nebulisers consist of theactive ingredient in a liquid carrier and comprising up to 40% w/w ofthe formulation, preferably less than 20% w/w. The carrier is typicallywater or a dilute aqueous alcoholic solution, preferably made isotonicwith body fluids by the addition of, for example, sodium chloride.Optional additives include preservatives if the formulation is notprepared sterile, for example, methyl hydroxy-benzoate, anti-oxidants,flavouring agents, volatile oils, buffering agents and surfactants.

Suitable formulations for administration by insufflation include finelycomminuted powders which may be delivered by means of an insufflator ortaken into the nasal cavity in the manner of a snuff. In theinsufflator, the powder is contained in capsules or cartridges,typically made of gelatin or plastic, which are either pierced or openedin situ and the powder delivered by air drawn through the device uponinhalation or by means of a manually-operated pump. The powder employedin the insufflator consists either solely of the active ingredient or ofa powder blend comprising the active ingredient, a suitable powderdiluent, such as lactose, and an optional surfactant. The activeingredient typically comprises from 0.1 to 100 w/w of the formulation.

Therefore, according to a further aspect of the present invention, thereis provided the use of a compound of formula (I) or a pharmaceuticallyacceptable solvate thereof in the preparation of a medicament for theprophylaxis or treatment of an inflammatory condition or immunedisorder.

Compounds according to the invention can be made according to anysuitable method of organic chemistry. Therefore, according to a furtheraspect of the invention, there is provided a process for preparing thecompounds of formula (I), or solvates thereof which comprises reactingthe compound of formula (II)

or an activated derivative thereof with a compound of formula (III)

wherein Q, X, R¹, R, R² and R³ and n are as hereinbefore defined.

and optionally converting the compound of formula (I) so formed to adifferent compound of formula (I) or to a corresponding solvate.

When X is oxygen, the esterification may be effected by standardmethods, for example using an acid catalyst and, optionally, an inertsolvent such as toluene, benzene, or a xylene. Suitable acid catalystsinclude mineral acids; for example, sulphuric acid, hydrochloric acid,and phosporic acid; and organic acids; for example, methanesulphonicacid, or toluenesulphonic acid. The esterification is typically carriedout at elevated temperature, for example, 50-150° C., preferably withremoval of the water formed by distillation.

Where X is oxygen or —NH—, the reaction may be effected by firstpreparing an activated derivative of the compound of formula (II).Suitable activated derivatives include activated esters or acid halidesand may either be isolated before reaction with the compound of formula(III) or prepared in situ. Particularly useful activated esters of thecompound of formula (II) are acylimidazoles which are readily preparedby reaction of the compound of formula (II) with N,N¹-carbonyldiimidazole.

Conversion of an activated derivative of the compound of formula (II) toa compound of formula (I) may be effected in an inert solvent, optimallyin the presence of a non-nucleophilic base, such as potassiumt-butoxide, sodium hydride, or a non-nucleophilic organic base, such as1,8-diazabicylo [5.4.0] undec-7-ene.

The compound of formula (II) may be prepared as described in PCTapplication No. GB 9501808.

Compounds of formula (III) are commercially available or may be preparedby literature methods. For example, R. A. Bartsch et al, J. Org. Chem.1989, 54: 857-860 and J. M. Harris, Macromol. J. Sci. Rev. Polymer Phys.Chem. 1985, C25 (3): 325-373, and S. Zalopsky, Bioconjugate Chem. 1995,6: 150-165.

Alternatively, compounds of formula (I) may be prepared by condensationof a compound of formula (IV)

or an acetal derivative thereof,

wherein Q, X, R¹, n and R are as hereinbefore defined,

with 1,3-bis(cyclohexylmethyl)-5,6-diaminouracil (which may be preparedas described in the Examples). The condensation is suitably carried outin a polar solvent at non-extreme temperature as described in PCTApplication No. GB9501808.

Compounds of formula (IV) may be prepared by coupling a compound offormula (III) with the appropriate carboxylic acid. Methods foreffecting this coupling and for preparing the carboxylic acid aredescribed in PCT Application No. GB9501808.

Conversion of a compound of formula (I) to a solvate thereof may beeffected by standard methods known to a person skilled in the art.

The invention will now be described by way of illustration only, by thefollowing examples:

Reference Example EXAMPLE 1(E)-4-[1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]cinnamicacid (a) 1,3-Bis(cyclohexylmethyl)urea

A mixture of cyclohexanemethylamine (Aldrich, 68.66 g) and 5 N sodiumhydroxide (Fisher, 200 ml) was stirred vigorously with cooling (−10° C.)while a solution of phosgene (30.0 g) in toluene (600 ml) was addedrapidly. After stirring for 20 minutes, the resulting mixture wasfiltered and the precipitated solid was washed with water (˜1500 ml) anddried at 67 Pa (0.5 Torr) to give 1,3-bis(cyclohexylmethyl)urea as whitepowder (72.72 g, 95%), m.p. 150-152° C.; ¹H-NMR (DMSO-d₆) δ: 5.74 (br t,J=5.8 Hz, 2, 2 NH), 2.81 (t, J=6.3 Hz, 4, 2 NCH₂), 1.62, 1.25, and 0.85(all m, 22, 2 cyclohexyl).

Anal. Calcd for C₁₅H₂₈N₂O: C, 71.38; H, 11.18; N, 11.10. Found: C,71.22; H, 11.17; N, 11.15.

(b) 6-Amino-1,3-bis(cyclohexylmethyl)uracil

Cyanoacetic acid (Aldrich, 21.0 g) was dissolved in acetic anhydride(260 ml). This solution was added to 1,3-bis(cyclohexylmethyl)urea (fromstep (a), 54.5 g) and the solution maintained at 80° C. for 2 h undernitrogen. Volatiles were removed in vacuo and the residual oil dried byevaporation of portions of 10% water-ethanol (3×400 ml). The residualsolids were dissolved in ethanol (600 ml)-water(300 ml) at 80° C. withadjustment of the pH to 10 by addition of 10% aqueous sodium carbonate.The hot solution was diluted with water (75 ml) and cooled to ambienttemperature. The colorless crystals which formed were filtered off,washed with water (3×500 ml) and dried at 67 Pa (0.5 Torr) to give6-amino-1,3-bis(cyclohexylmethyl)uracil (64.98 g, 94%), m.p. 138-141°C.; ¹H-NMR (DMSO-d₆) δ: 6.73 (br s, 2, NH₂), 4.63 (s, 1, H-5), 3.67 (d,J=7.3 Hz, 2, NCH₂), 3.57 (d, J=7.3 Hz, 2, NCH₂), 1.55 and 1.09 (both m,22, 2 cyclohexyl).

Anal. Calcd for C₁₈H₂₉N₃O₂.H₂O: C, 64.07; H, 9.26; N, 12.45. Found: C,63.98; H, 9.27; N, 12.48.

6-Amino-1,3-bis(cyclohexylmethyl)-5-nitrosouracil

6-Amino-1,3-bis(cyclohexylmethyl)uracil (from step (b), 25.0 g) wasdissolved in glacial acetic acid (440 ml), water (440 ml) and ethanol(440 ml) at reflux. To this solution was added sodium nitrite (5.65 g).The resulting mixture was stirred as it cooled slowly to ambienttemperature. The lavender precipitate was filtered off, washed with 1:1water-ethanol and dried to give6-amino-1,3-bis(cyclohexylmethyl)-5-nitrosouracil as light purplecrystals (23.46 g, 86%), m.p. 240-243° C. dec with effervescence; ¹H-NMR(DMSQ-d₆) δ: 13.23 (br s, 1, =NOH), 9.00 (br s, 1, =NH), 3.73 (br t,J=6.86, 4, 2 NCH₂), 2.0-1.6 and 1.7-1.1 (both m, total 22, 2cyclohexyl).

Anal. Calcd for C₁₈H₂₈N₄O₃: C, 62.05; H, 8.10; N, 16.08. Found: C,62.13; H, 8.12; N, 16.03.

(d)(E)-4-[1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro2,6dioxo-9H-purin-8-yl]cinnamicacid

The title compound was prepared from1,3-bis(cyclohexylmethyl)-5,6-diaminouracil by the method of J.Perutmattam, Syn. Commun. 1989, 19:3367-3370.1,3-Bis(cyclohexylmethyl)-5,6-diaminouracil was freshly prepared byshaking a mixture of 6-amino 1,3-bis(cyclohexylmethyl)-5-nitrosouracil(from step (c), 5.00 g) in methanol (250 ml)-water (25 ml) with 10%palladium on carbon (0.50 g) under hydrogen at 344.7 kPa (50 psi) on aParr shaker for 2 h. The catalyst was filtered off (Celite™) and thecolorless filtrate was concentrated to 25 ml. 4-Formylcinnamic acid(Aldrich, 2.53 g, 14.35 mmol) was added to this solution of1,3-bis(cyclohexylmethyl)-5,6-diaminouracil and the resulting yellowmixture was concentrated and the residual yellow solid dried byevaporation of several portions of absolute ethanol. The resultingyellow powder (Schiff base intermediate) was stirred in dimethoxyethane(115 ml) with iodine (4.0 g) at 60° C. (oil bath) for 20 h. A saturatedaqueous solution of sodium thiosulfate was added to the warm reactionmixture until complete decolorization of iodine resulted. The paleyellow precipitate was filtered off, washed with water, and dried at 67Pa (0.5 Torr) to give (E)-4-[1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]cinnamicacid as a pale yellow powder (6.73 g, 91%), m.p. >300° C. Such sampleswere further purified by dissolving them in 1N aqueous sodium hydroxide,filtering the resulting hazy solution through Celite™, and acidifyingthe clear filtrate with hydrochloric acid. The resulting precipitate wasfiltered and washed with water to give title compound as a pale yellowpowder, m.p. >300° C.; ¹H-NMR (DMSO-d₆) δ: 13.80 and 12.40 (both br m, 1each, CO₂H and NH), 8.12 (d, J=8.3 Hz, 2, 2 phenyl CH), 7.84 (d, J=8.4Hz, 2, 2 phenyl CH), 7.64 (d, J=16.0 Hz, 1, CH=), 6.64 (d, J=16.0 Hz, 1,CH=), 3.93 (d, J=7.0 Hz, 2, CH₂N), 3.79 (d, J=6.8 Hz, 2, CH₂N), 2.0-1.4and 1.3-0.85 (both br m, 22 total, 2 cyclohexyl).

Anal. Calcd for C₂₈H₃₄N₄O₄: C, 68.55; H, 6.99; N, 11.42. Found: C,68.45; H, 6.98; N, 11.48.

Synthetic Examples EXAMPLE 2(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Triethylene Glycol Methyl Ether Ester

Triethylene glycol monomethyl ether (Aldrich, 80.0 g) was dried byevaporation of portions of xylenes (3×50 ml) under a stream of N₂ at125° C.(E)-4-[1,3Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6dioxo-9H-purin-8-yl]cinnamicacid (from step (d) example 1, 4.00 g) was then added to the glycol, andthe mixture was further dried by evaporation of xylenes (40 ml).Sulfuric acid (0.41 g) was added to the reaction mixture, which was thenheated to 190° C. Xylenes were added in 50 ml portions to replace thatbeing distilled off. After 2 h, the reaction mixture was treated withadditional sulfuric acid (0.2 g). After an additional 3 h at 140° C.,during which time the xylenes were continuously replaced, the reactionmixture was allowed to cool to ambient temperature, which caused a greatdeal of solid to precipitate from the brown solution. The mixture wasdiluted with chloroform (200 ml), and washed with water (4×50 ml). Theorganic layer was dried (sodium sulfate) and concentrated to afford ayellow solid, which was chromatographed on silica gel. The titlecompound eluted with 1-4% methanol in ethyl acetate, and wasrecrystallized from ethyl acetate by the addition of hexanes to providetitle compound as a white powder (3.2 g, 62%), m.p. 189 -192° C.; ¹H-NMR(DMSO-d₆) δ: 8.16 (d, J=8.0 Hz, 2, 2 phenyl CH), 7.88 (d, J=8.3 Hz, 2, 2phenyl CH), 7.70 (d, J=16.1 Hz, 1, CH=), 6.77 (d, J=16.1 Hz, 1, CH=),4.28 (m, 2, CO₂CH₂), 3.92 (d, J=6.8 Hz, 2, CH₂N), 3.78 (d, J=6.8 Hz, 2,CH₂N), 3.68 (m, 2, CH₂O), 3.6-3.5 (m, 6, 3 CH₂O), 3.40 (2, CH₂O), 3.23(s, 3, CH₃), 2.0-1.5 and 1.3-0.9 (both br m, 22 total, 2 cyclohexyl).

Anal. Calcd for C₃₅H₄₈N₄O₇: C, 66.02; H, 7.60; N, 8.80 Found: C, 65.91;H, 7.58; N, 8.76.

EXAMPLE 3(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Polyethylene Glycol (n=7.2) Methyl Ether Ester

A slurry of (E)-4-[1,3bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]cinnamicacid (from step (d) example 1, 0.50 g) in poly(ethyleneglycol)monomethyl ether (Aldrich, average molecular weight 350, 21 g, dried byevaporation of toluene prior to use) containing sulfuric acid (51 mg)was stirred at 133 Pa (1 Torr) for 15 min. The yellow reaction mixturewas then stirred at 190° C. (oilbath) and 133 Pa (1 Torr) for 3 h,during which time the solids dissolved, leaving a brown solution. Aftercooling to ambient temperature, the dark solution was poured onto water(100 ml). The aqueous mixture was stirred for 1.75 h before beingextracted with dichloroethane (3×30 ml). The combined extracts were thendried (sodium sulfate), and concentrated to a waxy solid, which waschromatographed on C-18 reverse phase silica gel (EM SeparationsLiChroprep RP-18). The column was eluted with a gradient from 10%water-methanol to neat methanol; crude product eluted in neat methanolas a yellow waxy solid, which was slurried in water (25 ml). The waterwas then evaporated under vacuum to provide the title compound as yellowwaxy solid (620 mg, 61%), m.p. 147-154° C.; ¹H-NMR (DMSO-d₆) δ: 8.15 (d,J=8.1 Hz, 2, 2 phenyl CH), 7.88 (d, J=8.2 Hz, 2, 2 phenyl CH), 7.70 (d,J=16.0 Hz, 1, CH=), 6.77 (d, J=16.0 Hz, 1, CH=), 4.28 (m, 2, CO₂CH₂),3.91 (d, J=7.0 Hz, 2, CH₂N), 3.78 (d, J=7.1 Hz, 2, CH₂N), 3.68 (m, 2,CH₂O), 3.6-3.38 (m, ca 25, ca 12.5 CH₂O), 3.22 (s, 3, CH₃), 2.0-1.5 and1.3-0.9 (both br m, 22 total, 2 cyclohexyl).

Anal. Calcd for C₂₉H₃₆N₄O₄ (C₂H₄O)_(7.2).0.6 H₂O: C, 62.56 H, 8.00; N,6.69. Found: C, 62.62; H, 8.01; N, 6.69.

EXAMPLE 4(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Tetraethylene Glycol Methyl Ether Ester

(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6dioxo-9H-purin-8-yl)cinnamicacid polyethylene glycol (n=7.2) methyl ether ester (from example 3, 2.0g) was separated into its components via repeated chromatographyemploying a Chromatotron (Harrison Research). Portions of the estermixture (250-350 mg) in ethyl acetate were applied to 1 mm thick silicaplates that had been preequilibrated with hexanes. The plates were theneluted with a gradient from 5-20% ethyl acetate in hexanes. Fractionscontaining discrete oligomers were isolated separately, and identicalfractions from the various plates were pooled and concentrated. Allmixed fractions were combined and rechromatographed. In this manner(E)-4-(1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicacid tetraethylene glycol methyl ether ester was obtained as a whitepowder (43 mg), m.p. 171-174° C.; ¹H-NMR (DMSO-d₆) δ: 8.18 (d, J=8.4 Hz,2, 2 phenyl CH), 7.91 (d, J=8.4 Hz, 2, 2 phenyl CH), 7.72 (d, J=16.1 Hz,1, CH=), 6.80 (d, J=16.1 Hz, 1, CH=), 4.30 (m, 2, CO₂CH₂), 3.94 (d,J=7.1 Hz, 2, CH₂N), 3.80 (d, J=7.1 Hz, 2, CH₂N), 3.71 (m, 2, CH₂O),3.58-3.42 (m, 12, 6 CH₂O), 3.24 (s, 3, CH₃), 2.0-1.5 and 1.3-0.9 (bothbr m, 22 total, 2 cyclohexyl).

Anal. Calcd for C₂₉H₃₆N₄O₄ (C₂H₄O)₄.0.6 H₂O: C, 64.25; H, 7.75; N, 8.10Found: C, 64.11; H, 7.56; N, 8.07.

EXAMPLE 5(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Pentaethylene Glycol Methyl Ether Ester

Chromatographic separation of(E)-4-(1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicacid polyethylene glycol (n=7.2) methyl ether ester (from example 3, 2.0g) into its components as described in example 4 provided the titlecompound as a yellow waxy solid (92 mg), m.p. 166-167° C.; ¹H-NMR(DMSO-d₆) δ: 8.18 (d, J=8.2 Hz, 2, 2 phenyl CH), 7.91 (d, J=8.2 Hz, 2, 2phenyl CH), 7.72 (d, J=16.0 Hz, 1, CH=), 6.79 (d, J=16.1 Hz, 1, CH=),4.30 (m, 2, CO₂CH₂), 3.94 (d, J=7.0 Hz, 2, CH₂N), 3.80 (d, J=7.0 Hz, 2,CH₂N), 3.70 (m, 2, CH₂O), 3.60-3.40 (m, 16, 8 CH₂O), 3.24 (s, 3, CH₃),2.0-1.5 and 1.3-0.9 (both br m, 22 total, 2 cyclohexyl).

Anal. Calcd for C₂₉H₃₆N₄O₄ (C₂H₄O)₅.0.15 H₂O: C, 64.38; H, 7.80; N, 7.70Found: C, 64.44; H, 7.90; N, 7.57.

EXAMPLE 6(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Hexaethylene Glycol Methyl Ether Ester

Chromatographic separation of(E)-4-(1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicacid polyethylene glycol (n=7.2) methyl ether ester (from example 3, 2.0g) into its components as described in example 4 provided the titlecompound as a yellow waxy solid (170 mg), m.p. 160-162° C.; ¹H-NMR(DMSO-d₆) δ: 8.18 (d, J=8.2 Hz, 2, 2 phenyl CH), 7.91 (d, J=8.2 Hz, 2, 2phenyl CH), 7.74 (d, J=16.0 Hz, 1, CH=), 6.79 (d, J=16.1 Hz, 1, CH=),4.30 (m, 2, CO₂CH₂), 3.93 (d, J=6.8 Hz, 2, CH₂N), 3.80 (d, J=7.2 Hz, 2,CH₂N), 3.70 (m, 2, CH₂O), 3.60-3.40 (m, 20, 10 CH₂O), 3.24 (s, 3, CH₃),2.0-1.5 and 1.3-0.9 (both br m, 22 total, 2 cyclohexyl).

Anal. Calcd for C₂₉H₃₆N₄O₄ (C₂H₄)₆.0.20 H₂O: C, 63.74; H, 7.88; N, 7.25Found: C, 63.69; H, 7.92; N, 7.34.

EXAMPLE 7(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Heptaethylene Glycol Methyl Ether Ester

Chromatographic separation of(E)-4-(1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicacid polyethylene glycol (n=7.2) methyl ether ester (from example 3, 2.0g) into its components as described in example 4 provided the titlecompound as a yellow waxy solid (105 mg), m.p. 154-156° C.; ¹H-NMR(DMSO-d₆) δ: 8.18 (d, J=8.4 Hz, 2, 2 phenyl CH), 7.91 (d, J=8.4 Hz, 2, 2phenyl CH), 7.73 (d, J=16.0 Hz, 1, CH=), 6.79 (d, J=16.1 Hz, 1, CH=),4.30 (m, 2, CO₂CH₂), 3.94 (d, J=7.0 Hz, 2, CH₂N), 3.80 (d, J=7.2 Hz, 2,CH₂N), 3.70 (m, 2, CH₂O), 3.60-3.40 (m, 24, 12 CH₂O), 3.24 (s, 3, CH₃),2.0-1.5 and 1.3-0.9 (both br m, 22 total, 2 cyclohexyl).

Anal. Calcd for C₂₉H₃₆N₄O₄ (C₂H₄O)₇.0.25 H₂O: C, 63.18; H, 7.95; N, 6.85Found: C, 63.15; H, 7.97; N, 6.93.

EXAMPLE 8(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Octaethylene Glycol Methyl Ether Ester

Chromatographic separation of(E)-4-(1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicacid polyethylene glycol (n=7.2) methyl ether ester (from example 3, 2.0g) into its components as described in example 4 provided the titlecompound as a yellow waxy solid (120 mg), m.p. 150-151° C.; ¹H-NMR(DMSO-d₆) δ: 8.18 (d, J=8.4 Hz, 2, 2 phenyl CH), 7.91 (d, J=8.6 Hz, 2, 2phenyl CH), 7.73 (d, J=16.0 Hz, 1, CH=), 6.79 (d, J=16.1 Hz, 1, CH=),4.30 (m, 2, CO₂CH₂), 3.94 (d, J=7.0 Hz, 2, CH₂N), 3.80 (d, J=7.2 Hz, 2,CH₂N), 3.70 (m, 2, CH₂O), 3.60-3.40 (m, 28, 14 CH₂O), 3.24 (s, 3, CH₃),2.0-1.5 and 1.3-0.9 (both br m, 22 total, 2 cyclohexyl).

Anal. Calcd for C₂₉H₃₆N₄O₄ (C₂H₄O)₈.0.25 H₂O: C, 62.73; H, 8.01; N, 6.50Found: C, 62.65; H, 8.10; N, 6.56.

EXAMPLE 9(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Decaethylene Glycol Methyl Ether Ester (a) Decaethylene GlycolMonomethyl Ether

Potassium t-butoxide (Aldrich, 95%, 239.24 g) was added in portions overa 1.25 h period to a solution of triethylene glycol monomethyl ether(Aldrich, 300 ml, 1.9 mol) and 1,2-bis(2-chloroethoxy)ethane (Aldrich,500 g). The reaction temperature was maintained at 16-20° C. during theaddition (icebath). After removal of the cooling bath, the reactiontemperature reached 30° C. before cooling to ambient temperature. Thereaction mixture was stirred at ambient temperature for 2 h, followed by18 h at 110° C., before volatiles were removed (2.5 kPa (19 Torr),110-125° C.). The viscous residue was then diluted with toluene (1.7 L),and filtered through Celite™. Toluene was distilled off such that thepot temperature never exceeded 165° C., and methyl hexaethylene glycolchloride was then isolated via fractional distillation of the lightbrown residue (80 Pa (0.6 Torr), 155-190° C., 95.78 g, 16%).

Potassium t-butoxide (Aldrich, 95%, 39.0 g) was added to a solution oftetraethylene glycol (Aldrich, 412.7 g) and methyl hexaethylene glycolchloride from above (95.8 g) at 18° C. over a 25 min period (ice/acetonebath). The reaction mixture was then stirred at 120° C. overnight. ThepH was adjusted to 7 by the addition of hydrochloric acid (12 N, 11.7ml), and volatiles were removed (64 Pa (0.48 Torr), up to 185° C.). Theresidual dark oil was diluted with toluene (250 ml), and treated withcalcium chloride (38.1 g). After stirring for 18 h, the mixture wasfiltered through Celite™ and concentrated to a dark oil (102 g), whichwas fractionally distilled to provide decaethylene glycol monomethylether as an amber oil (64.4 g, 45%). An analytical sample was obtainedvia chromatography on silica gel, eluting with 4% methanol in chloroformto provide a colorless oil; ¹H-NMR (DMSO-d₆) δ: 4.58 (t, J=5.5 Hz, 1,OH), 3.58-3.38 (m, 40, 20 OCH₂), 3.24 (s, 3, OCH₃).

Anal. Calcd for C₂₁H₄₄O₁₁: C, 53.37; H, 9.38. Found: C, 53.09; H 9.47.

(b) (E)-Methyl 4-(dimethoxymethyl)cinnamate

4-Formylcinnamic acid dimethyl acetal (Cleeland, Jr., et al., U.S. Pat.No. 3,969,373) (20.00 g) and anhydrous potassium carbonate (12.44 g)were stirred in anhydrous N,N-dimethylformamide (189 ml) for 5 minutes.Methyl iodide (12.8 g) was added and the resulting mixture was stirredvigorously with gentle heating (oil bath at 40° C.) for 18 h. Volatileswere evaporated in vacuo and the residue partitioned between hexanes(400 ml) and water (100 ml). The hexanes layer was dried (magnesiumsulfate) and evaporated to give (E)-methyl 4-(dimethoxymethyl)cinnamateas a pale yellow oil (18.98 g, 89%), ¹H-NMR (DMSO-d₆) consistent withstructure.

Anal. Calcd for C₁₃H₁₆O₄: C, 66.09; H, 6.83. Found: C, 65.96; H, 6.86.

(c) 4-Formylcinnamic Acid Decaethylene Glycol Methyl Ether Ester

A solution of methyl 4-(dimethoxymethyl)cinnamate (from step (b), 4.96g), decaethylene glycol monomethyl ether (from step (a), 14.87 g), andtitanium (IV) isopropoxide (Aldrich, 1.05 ml) was stirred at 110° C.under high vacuum for 18 h. The resulting black oil was then cooled to35° C., treated with hydrochloric acid (1 N, 24.5 ml), and extractedwith toluene (3×100 ml). The combined extracts were concentrated to adark oil, which was chromatographed on silica gel. The title compoundeluted with 10% methanol in chloroform as a yellow oil (4.60 g, 34%),¹H-NMR (DMSO-d₆) δ: 10.54 (s, 1, CHO), 7.98 (m, 4, 4 phenyl CH), 7.76(d, J=16 Hz, 1, CH=), 6.88 (d, J=16 Hz, 1, CH=), 4.31 (m, 2, CO₂CH₂),3.70 (m, 2, OCH₂), 3.51 (m, 36, 18 OCH₂) 3.25 (s, 3, OCH₃).

Anal. Calcd for C₃₁H₅₀O₁₃.0.65 H₂O: C, 537.96; H, 8.05. Found: C, 57.95;H, 7.95.

(d)(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Decaethylene Glycol Methyl Ether Ester

The title compound was prepared from1,3-bis(cyclohexylmethyl)-5,6-diaminouracil by the method of J.Perutmattam, Syn. Commun. 1989, 19:3367-3370. In the manner of step (d)of example 1, 6-amino 1,3-bis(cyclohexylmethyl)-5-nitrosouracil (fromstep (c) example 1, 2.00 g) was converted to1,3-bis(cyclohexylmethyl)-5,6-diaminouracil, which was then combinedwith 4-formylcinnamic acid decaethylene glycol monomethyl ether ester(from step (c), 3.62 g) in ethanol (50 ml). The resulting yellow mixturewas concentrated, and the residual yellow semi-solid was dried byevaporation of several portions of absolute ethanol. The resultingyellow semi-solid (Schiff base intermediate) was then stirred indimethoxyethane (60 ml) with iodine (1.60 g, 6.31 mmol) at 50° C.(oilbath) for 18 h. Sufficient saturated aqueous sodium thiosulfatesolution was added to the warm reaction mixture to effect completedecolorization of iodine. The aqueous mixture was concentrated to avolume of 20 ml, diluted with water (50 ml), and extracted withchloroform (4×50 ml). The combined organic layers were then dried(magnesium sulfate) and concentrated to give an oily solid, which waschromatographed on silica gel. The title compound eluted in 6% methanolin chloroform as a yellow oil (3.6 g), which was partitioned betweenchloroform (150 ml) and water (50 ml). The organic layer wasconcentrated, and the resulting oil was precipitated fromdichloromethane by the addition of hexanes to provide title compound asa yellow powder, which was then washed with hexanes and dried undervacuum at 56° C. (2.57 g, 47%), m.p. 143-145° C.; ¹H-NMR (DMSO-d₆) δ:8.16 (d, J=8.4 Hz, 2, 2 phenyl CH), 7.88 (d, J=8.5 Hz, 2, 2 phenyl CH),7.70 (d, J=16.0 Hz, 1, CH=), 6.78 (d, J=16.0 Hz, 1, CH=), 4.29 (m, 2,CO₂CH ₂), 3.92 (d, J=7.1 Hz, 2, CH₂N), 3.78 (d, J=7.1 Hz, 2, CH₂N), 3.69(t, J=4.6 Hz, 2, CH₂O), 3.60-3.35 (m, 36, 18 CH₂O), 3.23 (s, 3, CH₃),2.0-1.5 and 1.3-0.9 (both br m, 22 total, 2 cyclohexyl).

Anal. Calcd for C₄₉H₇₆N₄O₁₄: C, 62.28; H, 8.10; N, 5.93. Found: C,62.14; H, 8.06;N,6.02.

EXAMPLE 10 (E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic Acid NonaethyleneGlycol Methyl Ether Ester (a) Nonaethylene Glycol Monomethyl Ether

A mixture of hexaethylene glycol (Aldrich, 100 g) and benzyl bromide(Aldrich, 12 g) in aqueous sodium hydroxide (50% (w/w), 80 ml) wasstirred at 100° C. (oil bath) under nitrogen for 2 h. The reactionmixture was then cooled to ambient temperature, diluted with water to atotal volume of 500 ml, and extracted with diethyl ether (200 ml) toremove dibenzylated product. Sodium chloride (100 g) was added to theaqueous layer, which was extracted further with diethyl ether (6×100ml). These ether extracts were combined, dried (sodium sulfate), andconcentrated to give hexaethylene glycol monobenzyl ether as an oil (25g, 20% based on glycol), ¹H-NMR (CDCl₃) δ: 7.30 (m, 5, 5 phenyl CH),4.53 (s, 2, benzyl CH₂), 3.69-3.54 (m, 22, 11 OCH₂), 3.06 (br s, 3, OHand CH₂O).

A solution of toluene sulfonyl chloride (Aldrich, 38 g) and triethyleneglycol monomethyl ether (Aldrich, 16.4 g) in dry pyridine (150 ml) wasstirred at 0° C. (icebath) for 4 h, followed by 18 h at ambienttemperature. The solution was then poured onto ice water (500 ml) andextracted with diethyl ether (3×300 ml). The ether extracts werecombined, washed with hydrochloric acid (3 N) and water, dried (sodiumsulfate), and concentrated to provide triethylene glycol methyl tosylether as a colorless oil (20.0 g, 62% based on glycol), ¹H-NMR (CDCl₃)δ: 7.75 (d, J=8.0 Hz, 2, 2 phenyl CH), 7.30 (d, J=8.1 Hz, 2, 2 phenylCH), 4.11 (t, J=4.8 Hz, 2, CH₂OS), 3.65-3.41 (m, 10, 5 CH₂O), 3.32 (s,3, CH₃O) and 2.40 (s, 3, benzylic CH₃).

A solution of hexaethylene glycol monobenzyl ether from above (22.3 g)in anhydrous THF (100 ml) was added to a suspension of 50% NaH (3.5 g)in anhydrous THF (100 ml). The suspension was stirred at ambienttemperature for 30 min and then a solution of triethylene glycol methyltosyl ether from above (22.0 g) in THF (100 ml) was added dropwise. Themixture was refluxed under nitrogen overnight, cooled to ambienttemperature, quenched with water (500 ml), and extracted with diethylether (3×300 ml). The ether extracts were combined, dried (sodiumsulfate), and concentrated in vacuo to give nonaethylene glycol benzylmethyl ether as an oil (27 g, 88%), ¹H-NMR (CDCl₃) δ: 7.31 (m, 5, 5phenyl CH), 4.54 (s, 2, benzyl CH₂), 3.62-3.52 (m, 36, 18 CH₂O), 3.35(s,3, CH₃).

A solution of nonaethylene glycol benzyl methyl ether from above (38 g)in methanol (200 ml) was shaken with 10% palladium on activated charcal(Aldrich, 1.0 g) under hydrogen 344.7 kPa (50 psi) on a Parr apparatusovernight. The catalyst was filtered off (Celite™), and the filtrate wasconcentrated in vacuo to provide nonaethylene glycol monomethyl ether asan oil (23 g, 74%), ¹H-NMR (CDCl₃) δ: 3.67-3.47 (m, 36, 18 OCH₂), 3.32(s, 3, CH₃).

Anal. Calcd for C₁₉H₄₀O₁₀: C, 53.26; H, 9.41. Found: C, 53.25; H, 9.41.

(b)(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Nonaethylene Glycol Monomethyl Ether Ester

A mixture of 4-formylcinnamic acid (Aldrich, 22.0 g), nonaethyleneglycol monomethyl ether (from step (a), 60.0 g), and toluene sulfonicacid (Aldrich, 10 g) in dry xylenes (600 ml) was refluxed for 4 h(oilbath) until about 2.0 ml (110 mmol) of water had collected in a DeanStark trap. The reaction mixture was then concentrated to about 100 ml,cooled to ambient temperature, and passed through a silica gel column.4-Formylcinnamic acid nonaethylene glycol methyl ether ester eluted withchloroform:acetone (60:40) as an oil (72 g, 97%), ¹H-NMR (CDCl₃) d:10.01 (s,1, CHO), 7.89 (d, J=8.1 Hz, 2, 2 phenyl CH), 7.71 (d, J=16.1Hz, 1, CH=), 7.66 (d, J=8.0 Hz, 2, 2 phenyl CH), 6.57 (d, J=16.1 Hz, 1,CH=), 4.37 (m, 2, CO₂CH₂), 3.77 (m, 2, CH₂O), 3.67-3.52 (m, 32, 16CH₂O), 3.35 (s, 3, CH₃).

The title compound was prepared from1,3-bis(cyclohexylmethyl)-5,6-diaminouracil by the method of JPerutmattam, Syn. Commun. 1989, 19:3367-3370. In the manner of step (d)of example 9, 1,3-bis(cyclohexylmethyl)-5,6-diaminouracil (from step (d)example 1, 10.4 g) was condensed with 4-formylcinnamic acid nonaethyleneglycol monomethyl ether from above (18.0 g) to provide the titlecompound as a light yellow solid (20.0 g, 74%), m.p. 143-145° C.; ¹H-NMR(DMSO-d₆) δ: 8.28 (d, J=8.3 Hz, 2, 2 phenyl CH), 7.74 (d, J=16.2 Hz, 1,CH=), 7.67 (d, J=8.4 Hz, 2, 2 phenyl CH), 6.55 (d, J=15.9 Hz, 1, CH=),4.40 (m, 2, CO₂CH₂), 4.08 (d, J=6.9 Hz, 2, CH₂N), 4.00 (d, J=7.3 Hz, 2,CH₂N), 3.80 (m, 2, CH₂O), 3.72-3.52 (m, 32, 16 CH₂O), 3.35 (s, 3, CH₃),2.05-1.03 (m, 22, 2 cyclohexyl).

Anal. Calcd for C₄₇H₇₂N₄O₁₃: C, 62.65; H, 8.05; N, 6.32. Found: C,62.40; H, 7.92; N, 6.42.

EXAMPLE 11(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Polyethylene Glycol (n=11.7) Methyl Ether Ester

A slurry of (E)-4-[1,3bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]cinnamicacid (from step (d) example 1, 5.50 g) in poly(ethyleneglycol)monomethyl ether (Aldrich, average molecular weight 550, 365 g, dried byevaporation of toluene prior to use) containing sulfuric acid (0.57 g)was stirred at 133 Pa (1 Torr) for 15 min. The yellow reaction mixturewas then stirred at 190° C. (oilbath) and 133 Pa (1 Torr) for 3 h,during which time the solids dissolved, leaving a brown solution. Aftercooling to ambient temperature, the dark solution was poured onto water(150 ml). The aqueous mixture was stirred for 1.5 h before beingextracted with dichloroethane (3×120 ml). The combined extracts werethen washed with water (150 ml) and the pH of the aqueous layer wasadjusted to a value of 6.5 by the addition of concentrated ammoniumhydroxide. After drying (sodium sulfate), the extracts were concentratedand the resulting oil was chromatographed on C-18 reverse phase silicagel (EM Separations LiChroprep RP-18). The column was eluted with agradient from 30% water-methanol to neat methanol; crude product elutedin neat methanol. This orange waxy solid was further chromatographed onsilica gel. The title compound eluted in 1-5% methanol in chloroform asa yellow waxy solid, which was then dissolved in chloroform andprecipitated through the addition of hexanes to provide title compoundas a light-yellow waxy solid (6.34 g, 56%), m.p. 140-143° C.; ¹H-NMR(DMSO-d₆) δ: 8.15 (d, J=8.5 Hz, 2, 2 phenyl CH), 7.88 (d, J=8.5 Hz, 2, 2phenyl CH), 7.70 (d, J=16.0 Hz, 1, CH=) 6.77 (d, J=16.1 Hz, 1, CH=),4.28 (m, 2, CO₂CH₂), 3.92 (d, J=7.0 Hz, 2, CH₂N), 3.78 (d, J=17.2 Hz, 2,CH₂N), 3.70 (m, 2, CH₂O), 3.6-3.35 (m, ca 42, ca 10.5 CH₂CH₂O), 3.23 (s,3, CH₃), 2.0-1.5 and 1.3-0.9 (both br m, 22 total, 2 cyclohexyl).

Anal. Calcd for C₂₉H₃₆N₄O₄ (C₂H₄O)_(11.6).0.4 H₂O: C, 61.30; H, 8.20; N,5.48. Found: C, 61.24; H, 8.26; N, 5.54.

EXAMPLE 12 (E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic Acid HexaethyleneGlycol Ester (a) 4-Formylcinnamic Acid Hexaethylene Glycol Ester

(E)-Methyl 4-(dimethoxymethyl)cinnamate (from step (b) example 9, 16.1g, 68.1 mmol), and titanium isopropoxide (Aldrich, 6.34 g) were stirredin an excess of crude hexaethylene glycol mono(tetrahydropyranyl) ether(J. W. Cornforth, E. D. Morgan, K. T. Potts, R. J. W. Rees, Tetrahedron1973, 29:1659-1667, 52.76 g) at ambient temperature for 3 d. Thesolution was then stirred under vacuum 267 Pa (2 Torr) at 120 ° C. for 4h. After the solution had cooled to 32° C., hydrochloric acid (1 N, 80ml, 80 mmol) was added. The resulting aqueous solution was stirred for1.5 h at 32-40° C. before being diluted with water (10 ml). The solutionwas stirred for an additonal 30 min before another 10 ml portion ofwater was added. After 2.5 h, the solution was cooled to ambienttemperature and extracted with toluene (3×100 ml). The toluene extractswere combined, dried (sodium sulfate), and concentrated to an oil (33.81g), which was chromatographed on silica gel. The title compound elutedin 10% methanol in ethyl acetate, and was dried by evaporation ofethanol to provide title compound as a waxy yellow solid (8.80 g, 29%)¹H-NMR (DMSO-d₆) δ: 10.06 (s, 1, CHO), 7.98 (m, 4, 4 phenyl CH), 7.77(d, J=16.0 Hz, 1, CH=), 6.88 (d, J=16.0 Hz, 1, CH=), 4.60 (m, 1, OH),4.31 (m, 2, CO₂CH₂), 3.71 (m, 2, CH₂O), 3.51 (m, 20, 10 CH₂O).

Anal. Calcd for C₂₂H₃₂O₉.0.35 H₂O.0.25 C₂H₆O: C, 58.97; H, 7.52; Found:C, 58.43; H, 7.40.

(b)(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-Purin-8-yl)cinnamicAcid Hexaethylene Glycol Ester

The title compound was prepared from1,3-bis(cyclohexylmethyl)-5,6-diaminouracil by the method of J.Perutmattam, Syn. Commun. 1989, 19:3367-3370. In the manner of step (d)of Example 9, 1,3-bis(cyclohexylmethyl)-5,6-diaminouracil (from step (d)example 1, 7.68 g) was condensed with 4-formylcinnamic acid hexaethyleneglycol ester (from step (a), 10.52 g) to provide the title compound as alight yellow solid (8.0 g, 45%), m.p. 165-168° C.; ¹H-NMR (DMSO-d₆) δ:8.17 (d, J=8.4 Hz, 2, 2 phenyl CH), 7.90 (d, J=8.4 Hz, 2, 2 phenyl CH),7.78 (d, J=16.0 Hz, 1, CH=), 6.80 (d, J=16.0 Hz, 1, CH=), 4.60 (m, 1,OH), 4.30 (m, 2, CO₂CH₂), 3.93 (d, J=6.9 Hz, 2, CH₂N), 3.80 (d, J=7.2Hz, 2, CH₂N), 3.71 (m, 2, CH₂O), 3.65-3.40 (m, 20, 10 CH₂O), 2.1-1.5 and1.6-0.9 (both br m, 22 total, 2 cyclohexyl).

Anal. Calcd for C₄₀H₅₈N₄O₁₀.0.9 H₂O: C, 62.30; H, 7.82; N, 7.27 Found:C, 62.33; H, 7.80; N, 7.26.

EXAMPLE 13(E)-4-(1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicacid polyethylene glycol (n=23.9) methyl ether ester (a)(E)-1,3-Bis(cyclohexylmethyl)-8-(4-(2-(1H-imidazol-1-ylcarbonyl)vinyl)phenyl)-9H-purin-2,6(1H,3H)-dione

A slurry of (E)-4-[1,3bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]cinnamicacid (from step (d) example 1, 2.97 g) in anhydrousN,N-dimethylformamide (50 ml) was heated briefly to near reflux undernitrogen. N,N′-Carbonyldiimidazole (Lancaster Synthesis, 1.17 g) wasthen added to the pale yellow slurry, which thinned and turned orange asa gas evolved. Within minutes the slurry turned a bright yellow andthickened as a yellow solid formed. The mixture was stirred for 18 h,diluted with dichloromethane (30 ml), and filtered. The bright yellowfilter plug was washed with dichloromethane (30 ml), and partiallyair-dried. The wet solid was then dried at 13 Pa (0.1 Torr) and 40° C.to provide(E)-1,3-bis(cyclohexylmethyl)-8-(4-(2-(1H-imidazol-1-ylcarbonyl)vinyl)phenyl)-9H-purin-2,6(1H,3H)-dioneas a yellow powder (3.25 g, 96%), m.p. 310° C. (dec); ¹H-NMR (DMSO-d₆)δ: 8.74 (s, 1, imidazole CH), 8.20 (d, J=8.9 Hz, 2, 2 phenyl CH), 8.06(d, J=7.7 Hz, 2, 2 phenyl CH), 8.03 (d, J=14.8 Hz, 1, CH=), 7.93 (s, 1,imidazole CH), 7.72 (d, J=15.7 Hz, 1, CH=), 7.14 (s, 1, imidazole CH),3.92 (d, J=7.0 Hz, 2, CH₂N), 3.77 (d, J=7.4 Hz, 2, CH₂N), 2.00-1.50 and1.25-0.90 both br m, 22 total, 2 cyclohexyl).

Anal. Calcd for C₃₁H₃₆N₆O₃.0.35 C₃H₇NO: C, 67.98; H, 6.84; N, 15.71.Found: C, 67.93; H, 6.67; N, 15.92.

(b)(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Polyethylene Glycol (n=23.9) Methyl Ether Ester

A slurry of(E)-1,3-bis(cyclohexylmethyl)-8-(4-(2-(1H-imidazol-1-ylcarbonyl)vinyl)phenyl)-9H-purin-2,6(1H,3H)-dione(from step (a), 2.25 g) in anhydrous N,N-dimethyl formamide (50 ml) waswarmed to 60° C. (oil bath) under nitrogen. Poly(ethyleneglycol)monomethyl ether (Shearwater Polymers, average molecular weight 1100,4.80 g, dried by evaporation of toluene prior to use) was added to thebright yellow slurry, followed by 1,8-diazabicyclo[5.4.0]undec-7-ene(Aldrich, 658 ml). Addition of the base produced a deep red reactionmixture and caused almost complete dissolution of the acyl imidazole.The red mixture was stirred at 60° C. for 2.5 d, during which time allsolids dissolved. The pH of the light red solution was then adjusted toa value of 5 by the addition of sulfuric acid. Volatiles were removedfrom the resulting light yellow solution at 32 Pa (0.24 Torr) and 47° C.to provide a yellow oil (11.37 g), which was chromatographed on C-18reverse phase silica gel (EM Separations LiChroprep RP-18). The columnwas eluted with a gradient from 40% water-methanol to neat methanol;crude product eluted in neat methanol to afford a yellow solid (10.25g), which was further chromatographed on silica gel. The title compoundeluted with 4-10% methanol in dichloromethane as a pale yellow glass(5.73 g, 92%), which was triturated with hexanes to provide a waxyyellow solid, m.p. 97-98° C.; ¹H-NMR (DMSO-d₆) 67 : 8.18 (d, J=8.2 Hz,2, 2 phenyl CH), 7.91 ( d, J=8.6 Hz, 2, 2 phenyl CH), 7.72 (d, J=15.8Hz, 1, CH=), 6.79 (d, J=15.9 Hz, 1, CH=), 4.30 (m, 2, CO₂CH₂), 3.94 (d,J=7.0 Hz, 2, CH₂N), 3.80 (d, J=7.8 Hz, 2, CH₂N), 3.70 (m, 2, CH₂O),3.57-3.41 (m, ca 88, ca 44 CH₂O), 3.25 (s, 3, CH₃), 2.0-1.5 and 1.3-0.9(both br m, 22 total, 2 cyclo

Anal. Calcd for C₂₉H₃₆N₄O₄ (C₂H₄O)_(23.4).0.30 H₂O: C, 59.02; H, 8.53;N, 3.58. Found: C, 59.05; H, 8.57; N, 3.62.

EXAMPLE 14(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Polyethylene Glycol (n=41.5) Methyl Ether Ester

In the manner of example 13,(E)-1,3-bis(cyclohexylmethyl)-8-(4-(2-(1H-imidazol-1-ylcarbonyl)vinyl)phenyl)-9H-purin-2,6(1H,3H)-dione(from step (a) example 13, 2.16 g) was coupled to poly(ethyleneglycol)monomethyl ether (Aldrich, average molecular weight 2000, 9.60 g) toprovide the title compound as a yellow powder (6.80 g, 76%), m.p. 56-64°C.; ¹H-NMR (DMSO-d₆) 67 : 8.18 (d, J=8.2 Hz, 2, 2 phenyl CH), 7.91 (d,J=8.2 Hz, 2, 2 phenyl CH), 7.72 (d, J=16.0 Hz, 1, CH=), 6.80 (d, J=16.1Hz, 1, CH=), 4.30 (m, 2, CO₂CH₂), 3.70-4.00 (m, 6, 2 CH₂N, and CH₂O),3.70-3.40 (m, ca 160, ca 80 CH₂O), 3.25 (s, 3, CH₃), 2.0-1.5 and 1.3-0.9(both br m, 22 total, 2 cyc

Anal. Calcd for C29H36N₄O₄ (C₂H₄)_(41.5): C, 57.67; H, 8.73; N, 2.40.Found: C, 57.51; H, 8.51; N, 2.31.

EXAMPLE 15(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic Acid Polyethylene Glycol (n=15) Methyl Ether Ester

In the manner of example 13,(E)-1,3-bis(cyclohexylmethyl)-8-(4-(2-(1H-imidazol-1-ylcarbonyl)vinyl)phenyl)-9H-purin-2,6(1H,3H)-dione(from step (a) example 13, 2.16 g, 3.84 mmol) was coupled topoly(ethyleneglycol) monomethyl ether (Aldrich, average molecular weight750, 3.30 g, 4.40 mmol) to provide the title compound as a yellow waxysolid (3.30 g, 74%), m.p. 124-125° C.; ¹H-NMR (DMSO-d₆) δ: 8.18 (d,J=8.2 Hz, 2, 2 phenyl CH), 7.91 (d, J=8.4 Hz, 2, 2 phenyl CH), 7.72 (d,J=16.0 Hz, 1, CH=), 6.80 (d, J=16.0 Hz, 1, CH=), 4.31 (m, 2, CO₂CH₂),3.94 (m, 2, CH₂N), 3.80 (m, 2, CH₂N), 3.70 (m, 2, CH₂O), 3.6-3.4 (m, ca58, ca 29 CH₂O), 3.25 (s, 3, CH₃), 1.8-1.5 and 1.3-0.9 (both br m, 22total, 2 cyclohexyl).

Anal. Calcd for C₂₉H₃₆N₄O₄ (C₂H₄)₁₅.0.5 H₂O: C, 57.67; H, 8.73; N, 2.40.Found: C, 57.51; H, 8.51; N, 2.31.

EXAMPLE 16(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Polyethylene Glycol (n=32.2) Ester

(E)-1,3-Bis(cyclohexylmethyl)-8-(4-(2-( 1H-imidazol-1-ylcarbonyl)vinyl)phenyl)-9H-purin-2,6(1H,3H)-dione (fromstep (a) example 13, 1.69 g) was added to molten poly(ethyleneglycol)(Aldrich, average molecular weight 1500, 90.0 g, 60.0 mmol, dried byevaporation of toluene prior to use) under nitrogen. The yellow slurrywas diluted with anhydrous N,N-dimethyl formamide (40 ml), and heated to60° C. (oilbath). 1,8-Diazabicyclo[5.4.0]undec-7-ene (Aldrich, 494 ml)was then added, producing a deep red reaction mixture and causing almostcomplete dissolution of the acyl imidazole. The mixture was stirred at60° C. for 16.5 h, during which time all solids dissolved. The pH of theorange solution was then adjusted to 5 through the addition of sulfuricacid. Volatiles were removed from the resulting yellow solution at 93 Pa(0.7 Torr) and 50° C., and the residual orange oil was chromatographedon C-18 reverse phase silica gel (EM Separations LiChroprep RP-18). Thecolumn was eluted with a gradient from 40% water-methanol to neatmethanol; crude product eluted in neat methanol to give a yellow film(8.50 g), which was further chromatographed on silica gel. The titlecompound eluted with 6-15% methanol in dichloromethane as a pale yellowglass (5.83 g), which was triturated with hexanes to provide a waxyyellow solid (4.773 g, 83%), m.p. 83-84° C.; ¹H-NMR (DMSO-d₆) δ: 13.92(s, 1 NH), 8.14 (d, J=8.4 Hz, 2, 2 phenyl CH), 7.87 (d, J=8.5 Hz, 2, 2phenyl CH), 7.68 (d, J=15.7 Hz, 1, CH=), 6.76 (d, J=16.1 Hz, 1, CH=),4.56 (t, J=5.5 Hz, 1, OH), 4.26 (m, 2, CO₂CH₂), 3.90 (d, J=7.3 Hz, 2,CH₂N), 3.76 (d, J=6.9 Hz, 2, CH₂N), 3.66 (m, 2, CH₂O), 3.6-3.2 (m, ca120, ca 60 CH₂O), 2.0-1.5 and 1.3-0.8 (both br m, 22 total, 2cyclohexyl).

Anal. Calcd for C₂₈H₃₄N₄O₄ (C₂H₄O)_(32.2).0.2 H₂O: C, 58.02; H, 8.60; N,2.93 Found: C, 58.01; H, 8.59; N, 2.92.

EXAMPLE 17(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6dioxo-9H-purin-8-yl)cinnamicAcid Polyethylene Glycol (n=18.9) Ester

In the manner of example 16,(E)-1,3-bis(cyclohexylmethyl)-8-(4-(2-(1H-imidazol-1-ylcarbonyl)vinyl)phenyl)-9H-purin-2,6(1H,3H)-dione(from step (a) example 13, 1.69 g) was coupled to poly(ethyleneglycol)(Aldrich, average molecular weight 1000, 60.0 g) to provide the titlecompound as a yellow waxy solid (2.747 g, 85%), m.p. <40° C.; ¹H-NMR(DMSO-d₆) δ: 13.88 (s, 1, NH), 8.11 (d, J=8.0 Hz, 2, 2 phenyl CH), 7.83(d, J=8.4 Hz, 2, 2 phenyl CH), 7.66 (d, J=16.1 Hz, 1, CH=), 6.73 (d,J=16.1 Hz, 1, CH=), 4.55 (t, J=5.5 Hz, OH), 4.26 (m, 2, CO₂CH₂), 3.87(d, J=6.9 Hz, 2, CH₂N), 3.74 (d, J=7.0 Hz, 2, CH₂N), 3.66 (m, 2, CH₂O),3.6-3.3 (m, ca 88, ca 40 CH₂O, overlapping H₂O), 2.1-1.5 and 1.3-0.9(both br m, 22 total, 2 cyclohexyl).

Anal. Calcd for C₂₈H₃₄N₄O₄ (C₂H₄O)_(18.9).1.2 H₂O: C, 58.77; H, 8.39; N,4.17. Found: C, 58.77; H, 8.28; N, 4.10.

EXAMPLE 18(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Polyethylene Glycol (n=13) Ester

In the manner of example 16,(E)-1,3-bis(cyclohexylmethyl)-8-(4-(2-(1H-imidazol-1-ylcarbonyl)vinyl)phenyl)-9H-purin-2,6(1H,3H)-dione(from step (a) example 13, 5.553 g) was coupled to poly(ethyleneglycol)(Dow, average molecular weight 600, 121.0 g) to provide the titlecompound as a yellow waxy solid (6.94 g, 64%), m.p. 142-143° C.; ¹H-NMR(DMSO-d₆) δ: 8.18 (d, J=8.4 Hz, 2, 2 phenyl CH), 7.90 (d, J=8.4 Hz, 2, 2phenyl CH), 7.72 (d, J=16.0 Hz, 1, CH=), 6.79 (d, J=16.0 Hz, 1, CH=),4.58 (t, J=5.3 Hz, OH), 4.30 (m, 2, CO₂CH₂), 3.94 (d, J=7.2 Hz, 2,CH₂N), 3.80 (d, J=7.0 Hz, 2, CH₂N), 3.70 (m, 2, CH₂O), 3.61-3.42 (m, ca48, ca 24 CH₂O), 2.0-1.5 and 1.3-0.9 (both br m, 22 total, 2cyclohexyl).

Anal. Calcd for C₂₈H₃₄N₄O₄ (C₂H₄O)₁₃0.1 H₂O; C, 59.98; H, 8.20; N, 5.18Found: C, 59.96; H, 8.18; N, 5.13.

EXAMPLE 193-{4-[1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]phenyl}propionicAcid Nonaethylene Glycol Monomethyl Ether Ester

A solution of(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicacid nonaethylene glycol monomethyl ether ester (from step (b) example10, 200 mg) in isopropanol (50 ml) was stirred under H₂ at 2.0 kPa (0.02Bar) in the presence of 10% palladium on activated charcoal (Aldrich, 40mg) in a Büchi Pressflow Hydrogenator for 23 h. The catalyst wasfiltered off (Celite™), and volatiles were evaporated under vaccuum.Chloroform (5×10 ml) was evaporated from the oily residue, which, afterdrying (0.2 mm Hg, 50° C.) afforded the title compound as a waxy yellowsolid (165 mg, 83%), m.p. 85-86° C., ¹H-NMR (DMSO-d₆) consistent withstructure.

Anal. Calcd for C₄₇H₇₄N₄O₁₃: C, 62.51; H, 8.26; N, 6.21. Found: C,62.39; H, 8.27; N, 6.27.

EXAMPLE 203-[4-[1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]phenyl}propiolicAcid Nonaethylene Glycol Methyl Ether Ester (a)3-[4-[1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]phenyl}-2,3-dibromopropionicAcid Ethyl Ester

A solution of 0.64 g (4.0 mmol) bromine in 10 mL chloroform was added toa stirred solution of 1.90 g (3.66 mmol)(E)-4-(1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicacid ether ester in 25 mL chloroform. The mixture was stirred at roomtemperature, then the solvent was evaporated to yield the title compoundas a white solid. (2.5 g, 100%) ¹H-NMR (CDCl₃)δ: 8.31 (d, J=8.4 Hz,phenyl CH, 2H), 7.56 (d, J=8.4 Hz, phenyl CH, 2H), 5.40 (d, J=11.7 Hz,CHBr, 1H), 4.83 (d, J=11.7 Hz, CHBr, 1H), 4.38 (q, J=7.3 Hz, CO₂CH₂,2H), 4.08 (d, J=7.2 Hz, CH₂N, 2H), 4.03 (d, J=7.3 Hz, CH₂N, 2H), 2.04(m, c-hexyl CH, 1H), 1.89 (m, c-hexyl CH, 1H), 1.5-1.8 (m, c-hexyl CH₂,10 H), 1.39 (t, J=7.3 Hz, terminal-CH₃, 3H), 1.0-1.3 (m, c-hexyl CH₂, 10H).

(b)3-[4-[1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]phenyl}propiolicAcid

2.8 g (4.1 mmol)3-[4-[1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]phenyl}-2,3-dibromo-propionicacid ethyl ester (part a of this example) was added to 56 mL of 1Mpotassium t-butoxide in t-butanol (Aldrich) at room temperature. Themixture was stirred at room temperature for 2.5 hours, then 0.10 mLwater was added. The solution was stirred for an additional hour then 1g charcoal was added in 250 mL water. The mixture was stirred for 30minutes then filtered. The filtrate was acidified with conc.hydrochloric acid (pH=1), then the resulting suspension was stirred for15 minutes, filtered then washed with 3×50 mL water. The solid was driedunder reduced pressure to give 1.63 g (81%) of title compound; ¹H-NMR(DMSO-d₆)δ: 8.15 (d, J=8.0 Hz, phenyl CH, 2H), 7.73 (d, J=8.0 Hz, phenylCH, 2H), 3.87 (d, J=6.6 Hz, CH₂N, 2H), 3.74 (d, J=6.8 Hz, CH₂N, 2H),1.88 (br s, c-hexyl CH, 1H), 1.4-1.8 (m, c-hexyl CH, CH₂, 11H), 0.8-1.2(m, c-hexyl CH₂, 10 H); MS (ES⁻): 487 (M-1), 443 (M-1-CO₂).

(c)3-[4-[1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]phenyl}propiolicAcid Nonaethylene Glycol Methyl Ether Ester

A solution of 324 mg (2.0) mmol 1,1′-carbonyldiimidazole in 10 mLacetonitrile was added to a stirred solution of 650 mg (1.33 mmol)3-[4-[1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]phenyl}propiolicacid in 40 mL anhydrous tetrahydrofuran. The mixture was stirred for 4hours, then 40 mL acetonitrile was added, and the suspension was stirredfor an additional 30 minutes and filtered. The solid was washed with2×10 mL acetonitrile and dried overnight under reduced pressure, thenmixed with 10 mL of anhydrous dimethyl formamide. Nonaethylene glycolmonomethyl ether (857 mg) was added to the suspension and the mixturewas stirred under nitrogen. 1,8diazabicyclo[5.4.0]undec-7-ene (300 μL)was added dropwise and the red solution was stirred at room temperaturefor an hour, then at 40° C. for an additional hour. The solution wascooled to 20° C., 100 mL of methylene chloride added, and the pHadjusted to 5 with 1 M potassium hydrogensulfate. Phases were separatedand the aqueous phase was washed with 2×20 mL methylene chloride. Thecombined organic phases were dried with anhydrous magnesium sulfate,then filtered. The filtrate was concentrated under reduced pressure, andthe residue was purified by silica gel chromatography using ethylacetate/ethanol (9:1) as eluent, to obtain 105 mg (9%) title compound;¹H-NMR (CDCl₃) δ: 8.27 (d, J=8.3 Hz, phenyl CH, 2H), 7.71 (d, J=8.3 Hz,phenyl CH, 2H), 4.41 (t, J=4.6 Hz, CO₂CH₂, 2H), 4.06 (d, J=7.3 Hz, CH₂N,2H), 3.97 (d, J=7.2 Hz, CH₂N, 2H), 3.78 (t, J=4.8 Hz, CH₂O, 2H), 3.6-3.7(m, 15 CH₂, 30 H), 3.52 (m, CH₂O, 2H), 3.34 (s, OCH₃, 3H), 2.04 (m,c-hexyl CH, 1H), 1.85 (m, c-hexyl CH, 1H), 1.6-1.8 (m, c-hexyl CH, CH₂,10H), 1.0-1.2 (m, c-hexyl CH₂, 10 H). MS (FAB⁺): 899 (M+1), 921 (M+Na).

EXAMPLE 21(E)-3-[1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]cinnamicAcid Nonaethylene Glycol Methyl Ether Ester (a)(E)-3-[(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]cinnamicacid

As in part (d) of Example, 1, 6-amino1,3-bis(cyclohexylmethyl)-5-nitrosouracil (from step (c) example 1, 2.79g, 8.00 mmol) was reduced to1,3-bis(cyclohexylmethyl)-5,6-diaminouracil, and condensed with3-formylcinnamic acid (T. Higa, A. J. Krubsack, J. Org. Chem. 1975, 40:3037-3045, 1.424 g) to give(E)-3-[1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]cinnamicacid as an off-white solid (1.947 g, 49%), m.p. >350° C.; ¹H-NMR(DMSO-d₆) consistent with structure.

Anal. Calcd for C₂₈H₃₄N₄O₄.0.10 H₂O: C, 68.30; H, 7.00; N, 11.38. Found:C, 68.33; H, 6.93; N, 11.34.

(b)(E)-3-[1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]cinnamicAcid Nonaethylene Glycol Methyl Ether Ester

A slurry of(E)-3-[1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]cinnamicacid (from step (a), 0.50 g) in anhydrous N,N-dimethylformamide (10 mL)was heated briefly to near reflux under nitrogen.N,N′-carbonyldiimidazole (Aldrich, 0.202 g, 1.22 mmol) was then added tothe pale yellow slurry, which thinned and turned orange as a gasevolved. Within minutes the slurry turned a bright yellow and thickenedas a yellow solid formed. The mixture was stirred for 18 h, diluted withdichloromethane (30 mL), and filtered. The bright yellow solid waswashed with dichloromethane (30 mL), and dried at 40° C. to provide(E)-1,3-bis(cyclohexylmethyl)-8-(3-(2-(1H-imidazol-1-ylcarbonyl)vinyl)phenyl)-9H-purin-2,6(1H,3H)-dioneas a yellow powder (0.403 g). To a mixture of this compound (0.40 g,0.74 mmol) and nonaethylene glycol monomethyl ether, (part (a) ofexample 10, 0.350 g) in N,N-dimethyl formamide (10 ml) was added1,8-diazabicyclo[5.4.0]undec-7-ene (Aldrich, 0.112 g). The resultingsolution was stirred at 55° C. for 20 h. The solution was cooled to roomtemperature and adjusted to pH 7 by addition of 1N HCl. Chloroform (50mL) was added and the solution was washed with water (2×20 mL). Thecombined organic layers were washed with brine, dried (magnesiumsulfate) and concentrated under reduced pressure. The residual waxysolid was chromatrographed on silica gel eluted with 10%methanol/chloroform to give(E)-3-[1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]cinnamicacid nonaethylene glycol methyl ether ester as a white waxy solid (0.421g, 63%); ¹H-NMR (DMSO-d₆)δ: 8.53 (s, 1, aryl CH), 8.13 (d, J=7.7, 1,aryl CH), and 7.80 (d, J=8.5, 1, aryl CH), 7.71 (d, J=16, 1, CH=) 7.58,(m, 1, aryl CH), 6.78, (d, J=15.9, 1, CH=) 4.28 (m,2, CH₂O),3.91, (d,J=7.2, 2,CH₂N), 3.77 (d, J=7.2, CH₂N), 3.68 (m, 2 CH₂O), 3.70-3.40 (m,32, 16 CH₂), 3.21 (s, 3, CH₃), 2.1-1.6 and 1.4-1.0 (m, 22, cyclohexylCH₂ and CH).

Anal. Calcd. For C₄₇H₇₂N₄O₁₃: C, 62.65; H, 8.05; N, 6.22. Found: C,62.57; H, 7.83; N, 6.50

EXAMPLE 22(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Nonaethylene Glycol Methyl Ether Amide (a) 2, 5, 8, 11, 14, 17, 20,23, 26-nonaoxo-octacosyl-28-amine

Sodium hydride (8.6 g, 344 mmol as 95%) was added to a solution ofhexaethylene glycol (Aldrich, 100 g) in anhydrous tetrahydrofuran (1000mL) at 15° C. The resulting mixture was stirred while coming to ambienttemperature over 1 h. Benzyl bromide (Aldrich, 59.9 g) was addeddropwise over 1 h and the resulting mixture stirred at ambienttemperature for 16 h. The cooled mixture was diluted with water (200mL)and extracted with diethyl ether (3×350 mL). The combined diethyl etherextracts were washed with water (2×100 mL). The combined aqueous layerswere saturated with sodium chloride and extracted with methylenechloride (4×400 mL). The combined methylene chloride layers were washedwith saturated sodium chloride (200 mL) and dried (magnesium sulfate).Removal of the volatiles under reduced pressure left hexaethylene glycolmonobenzyl ether (80.5 g, 64%); ¹H-NMR identical with that described inpart (a) of Example 10. A solution of hexaethylene glycol monobenzylether (80.0 g) in anhydrous THF (750 mL) was added to a suspension ofsodium hydride (95%, 5.4 g) in tetrahydrofuran. The resulting mixturewas stirred at ambient temperature for 30 min, and then a solution oftriethylene glycol methyl tosyl ether (prepared as described in part (a)of Example 1, 68.4 g) in THF (100 mL) was added dropwise. The mixturewas refluxed under nitrogen overnight. Additional sodium hydride (2.5 g)was added and reflux continued an additional 24 h. The mixture wascooled (ice bath), quenched with water (2 L), and extracted with diethylether (2×200 nmL). The aqueous layer was washed with methylene chloride(2×250 mL). The combined organic layers were dried (magnesium sulfate)and concentrated to a brown oil which was filtered through a silica gelpad washed with methylene chloride. Methylene chloride was evaporated toleave nonaethylene glycol benzyl methyl ether as an oil (63.1 g, 57%),¹H-NMR (DMSO-d₆) δ: 7.23 (m, 5, 5 phenyl CH), 4.38 (s, 2, benzyl CH₂),3.50-3.30 (m, 36, 18 CH₂O), 3.13 (s, 3, CH₃).

A solution of nonaethylene glycol benzyl methyl ether (10 g, 19.3 mmol)in ethanol (200 mL) was shaken with 10% palladium on activated charcal(Aldrich, 1.0 g) under hydrogen at 344.7 kPa (50 psi) on a Parrapparatus for 3 h. The catalyst was filtered off (Celite™), and thefiltrate was concentrated in vacuo and dried by evaporation of tolueneto provide nonaethylene glycol monomethyl ether as an oil (8.17 g, 99%),¹H-NMR (DMSO-d₆) δ: 4.56 (t, 1 OH), 3.60-3.35 (m, 36, 18 OCH₂),3.22 (s,3, CH₃).

To a solution of nonaethylene glycol monomethyl ether (2.0 g. 4.7 mmol)in pyridine (15 mL) at 0° C. was added toluenesulfonyl chloride (1.35g). After stiring at room temperature overnight, the mixture was cooledto 0° and adjusted to pH 2 by addition of 12N HCl. Water (200 mL) wasadded and the solution was washed with methylene chloride (3×50 mL). Thecombined organic layers were washed with brine, dried (magnesiumsulfate) and evaporated to provide nonaethylene glycol methyl tosylether as a colorless oil (2.7 g); ¹H-NMR (DMSO-d₆)δ: 7.85 and 7.55 (2d,4, C₆H₄), 4.18 (m, 2, CH₂OTos), 3.7-3.45 (m 34, 17 CH₂), 3.2 (s, 3,OCH₃) 2.40 (s, 3, CH₃).

To a solution of nonaethylene glycol methyl tosyl ether (2.6 g, 4.42mmol) in N,N-dimethylformamide (10 mL), was added sodium azide (Aldrich,0.35 g) and sodium iodide (Aldrich, 20 mg). The solution was refluxed 18h, cooled to room temperature, and diluted with chloroform (50 mL). Thissolution was washed with water (2×10 mL) and the organic layer dried(magnesium sulfate) and concentrated to a colorless oil (2.25 g). Theoil was dissolved in ethanol (100 mL), and stirred with 10% palladium oncarbon (Aldrich, 200 mg ) under hydrogen at 103.4 kPa (15 psi) for 48 hon a Buchi hydrogenation apparatus. The catalyst was removed byfiltration (Celite™) and solvent evaporated to leave 2, 5, 8, 11, 14,17, 20, 23, 26-nonaoxo-octacosyl-28-amine as a colorless oil (1.59 g,77%); ¹H-NMR (DMSO-d₆)δ: 3.60-3.40 (m, 36, 18 CH₂ and NH₂), 3.20 (3,CH₃); MS (CI) 428 (100%, M+1).

(b)(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Nonaethylene Glycol Methyl Ether Amide

To a mixture ofE)-1,3-bis(cyclohexylmethyl)-8-(4-(2-(1H-imidazol-1-ylcarbonyl)vinyl)phenyl)-9H-purin-2,6(1H,3H)-dione(part (a) of example 13), (0.5 g, 0.92 mmol) and the 2, 5, 8, 14, 17,20, 23, 26-nonaoxo-octacosyl-28-amine (part (a) of this example, 0.43 g,1.0 mmol) in N,N-dimethyl formamide (10 mL) was added1,8-diazabicyclo[5.4.0]undec-7-ene (Aldrich, 0.168 g). The resulting redsolution was stirred at 55° C. for 18 h. Additional amine (0.43 g) wasadded and heating continued for an additional 20 h. The resultingsolution was cooled to room temperature and adjusted to pH 7.0 byaddition of 1N hydrochloric acid. Water (25 mL) was added and thesolution was washed with ethyl acetate (50 mL). The organic layer waswashed with brine, dried (magnesium sulfate) and concentrated underreduced pressure. Chromatography on silica gel eluted the title compoundwith 10% methanol-chloroform. Evaporation of solvent left title compoundas a yellow solid (64 mg, 8%); ¹H-NMR (DMSO-d₆)δ: 8.24 (t, 1, NH), 8.15(d, J=8.4, 2, 2 aryl CH), 7.69 (d, J=8.3, 2, 2 aryl CH), 7.47 (d,J=15.5, 1, CH=), 6.75 (d, J=15.8, 1, CH=), 3.92 (d, J=7.1, 2, CH₂N),3.78 (d, J=7.2, 2, CH₂N), 3.6-3.3 (m, 36, 18 CH₂), 3.23 (s, 3, CH₃),2.0-1.5 and 1.25-0.95 (both m, 22, cyclohexyl CH₂ and CH).

Anal. Calcd for C₄₇H₇₃N₅O₁₂.0.85 H₂O: C, 61.67; H, 8.22; N, 7.65. Found:C, 61.66; H, 8.07; N, 7.67.

EXAMPLE 23(E)3-[(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]benzoicacid Nonaethylene Glycol Methyl Ether Ester (a)(E)-3-[(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]benzoicacid

1,3-Bis(cyclohexylmethyl)-5,6-diaminouracil was prepared as in part (d)of Example 1 by reduction of 1, 6-amino1,3-bis(cyclohexylmethyl)-5-nitrosouracil (2.00 g) and immediatelycondensed with 3-formylbenzoic acid (Aldrich, 1.424 g) by the method ofJ. Perumattam (Synthetic Commun. 1989, 19: 3367-3370) to give titlecompound as an off-white solid (2.27 g, 85%), m.p. >250° C.; ¹H-NMR(DMSO-d₆) consistent with structure.

Anal. Calcd for C₂₆H₃₂N₄O₄: C, 67.22; H, 6.94; N, 12.06. Found: C,67.10; H, 6.97; N, 12.04.

(b)(E)-3-[(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]benzoicacid Nonaethylene Glycol Methyl Ether Ester

A slurry of(E)-3-[(1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]benzoicacid (from part (a) of this example, 0.500 g) in anhydrousN,N-dimethylformamide (10 mL) was heated briefly to near reflux undernitrogen. N,N′-Carbonyldiimidazole (Aldrich, 0.213 g) was added to thepale yellow slurry, which thinned and turned orange as a gas evolved.Within minutes the slurry turned bright yellow and yellow solidprecipitated. The mixture was stirred for 18 h, diluted withdichloromethane (30 mL), and filtered. The solid was washed withdichloromethane (30 mL) and dried at 40° C. to provide(E)-1,3-bis(cyclohexylmethyl)-8-[3-(2-(1H-imidazol-1-ylcarbonyl)vinyl)phenyl]-9H-purin-2,6(1H,3H)-dioneas a yellow powder (0.46 g). A mixture of this solid (0.45 g),nonaethylene glycol monomethyl ether, (from part (a) of example 10,0.393 g) and anhydrous potassium carbonate (0.242 g) in acetonitrile (10mL) was stirred at reflux for 20 h. Chloroform (50 mL) was added and thesolution was washed with 1N HCl, (20 mL). The organic layer was washedwith brine, dried (magnesium sulfate), and chromatographed on silicagel. Elution with 10% methanol-chloroform evaporation of solvents gave(E)-3-[1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]benzoicacid nonaethylene glycol methyl ether ester as a waxy white solid (0.262g, 38%); ¹H-NMR (DMSO-d₆)δ: 8.79 (s) and 8.43 (d, J=7.9), 8.11 (d,J=8.0), and 7.57 (m, each 1,C₆H₄), 4.50 (m, 2, CH₂O), 3.99 (d, J=7.1, 2,CH₂N), 3.83 (m, 4, CH₂N and CH₂O), 3.70-3.40 (m, 32, 16 CH₂), 3.28 (s,3, CH₃), 2.1-1.6 and 1.4-1.0 (both m, 22, cyclohexyl CH₂ and CH).

Anal. Calcd. For C₄₅H₇₀N₄O₁₃.0.52 H₂O: C, 61.05; H, 8.10; N, 6.33.Found: C, 61.05; H, 8.09; N, 6.25.

EXAMPLE 24(E)-4-[(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]benzoicacid Nonaethylene Glycol Methyl Ether Ester (a)(E)-4-[(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]benzoicacid

1,3-Bis(cyclohexylmethyl)-5,6-diaminouracil was prepared as in part (d)of Example 1 by reduction of 1, 6-amino1,3-bis(cyclohexylmethyl)-5-nitrosouracil (1.00 g) and immediatelycondensed with 4-formylbenzoic acid (Aldrich, 1.424 g) by the method ofJ. Perumattam (Synthetic Commun. 1989, 19: 3367-3370) to give titlecompound as an off-white solid an off-white solid (720 mg, 54%),m.p. >300° C.; ¹H-NMR (DMSO-d₆) consistent with structure.

Anal. Calcd for C₂₆H₃₂N₄O₄: C, 67.23; H, 6.94; N, 12.06. Found: C,67.29; H, 6.98; N, 12.02.

(b)(E)-4-[(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]benzoicacid Nonaethylene Glycol Methyl Ether Ester

A slurry of(E)-4-[(1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]benzoicacid (from part (a) of this example, 0.50 g) in anhydrousN,N-dimethylformamide (10 mL) was heated briefly to near reflux undernitrogen. N,N′-Carbonyldiimidazole (Aldrich, 0.211 g) was then added tothe pale yellow slurry, which thinned and turned orange as a gasevolved. Within minutes the slurry turned a bright yellow and thickenedas a yellow solid formed. The mixture was stirred for 18 h, diluted withdichloromethane (30 nmL), and filtered. The bright yellow solid waswashed with dichloromethane (30 mL), and dried at 40° C. to provide(E)-1,3-bis(cyclohexylmethyl)-8-[3-(2-(1H-imidazol-1-ylcarbonyl)vinyl)phenyl]-9H-purin-2,6(1H,3H)-dioneas a yellow powder (0.32 g). A mixture of this sample (0.32 g),nonaethylene glycol monomethyl ether (from part (a) of example 10, 0.277g) and anhydrous potassium carbonate (0.170 g) in acetonitrile (10 mL)was refluxed for 20 h. Chloroform (50 mL) was added and the solution waswashed with 1N HCl, (20 mL). The organic layer was washed with brine,dried (magnesium sulfate), and eluted from a silica gel column with 10%methanol-chloroform. Evaporation of solvents left yellow waxy solidwhich was reprecipitated from ethyl acetate-hexanes. The yellow waxysolid precipitate was filtered and dried to(E)-4-[1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]benzoicacid nonaethylene glycol methyl ether ester (0.355 g, 65%); ¹H-NMR(DMSOd₆)δ: 8.24 (d, J=8.4, 2, 2 CH), 8.06 ( d, I=8.6, 2, 2 CH), 4.40(mn, 2, CH₂O), 3.90 (d, J=7.3, 2,CH₂N), 3.75 (m, 4, CH₂N and CH₂O),3.70-3.40 (m, 32, 16 CH₂), 3.19 (s, 3, CH₃), 2.1-1.6 and 1.4-1.0 (m, 22,cyclohexyl CH₂ and CH).

Anal. Calcd for C₄₅H₇₀N₄O₁₃: C, 61.77; H, 8.06; N, 6.40. Found: C,61.55; H, 7.99; N, 6.52

EXAMPLE 25(E)-2-[(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]benzoicacid Nonaethylene Glycol Methyl Ether Ester (a)(E)-2-[(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]benzoicacid

1,3-Bis(cyclohexylmethyl)-5,6-diaminouracil was prepared as in part (d)of Example 1 by reduction of 1, 6-amino1,3-bis(cyclohexylmethyl)-5-nitrosouracil (2.00 g) and immediatelycondensed with 2-formylbenzoic acid (Aldrich, 1.424 g) by the method ofJ. Perumattam (Synthetic Commun. 1989, 19: 3367-3370) to give titlecompound as an off-white solid off-white solid (1.22 g, 46%), m.p.271-274° C.; ¹H-NMR (DMSO-d₆) consistent with structure.

Anal. Calcd for C₂₆H₃₂N₄O₄: C, 67.22; H, 6.94; N, 12.06. Found: C,67.25; H, 6.99; N, 12.11.

(b)(E)-2-[(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]benzoicAcid Nonaethylene Glycol Methyl Ether Ester

(E)-2-[(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]benzoicacid (from part (a) of this example, 0.100 g) was converted by themethod of part (b) of Example 24 to(E)-2-[(1,3-bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl]benzoicacid nonaethylene glycol methyl ether ester (0.082 g, 43%), as a amberoil; ¹H-NMR (DMSO-d₆)δ:.7.80-7.50 (m, 4, 4 aromatic CH), 4.20 (m, 2,CH₂O), 3.80-3.60 (m, 4, 2CH₂N), 3.50-3.20 (m, 34, 17 CH₂), 3.20, (s,3,CH₃), 1.9-1.4 and 1.2-0.8 (m, 22, cyclohexyl).

Anal. Calcd. For C₄₅H₇₀N₄O₁₃.0.85 EtOAc 0.64 H₂O: C, 60.50; H, 8.18; N,5.83. Found: C, 60.50; H, 8.19; N, 5.83.

EXAMPLE 26(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-6-oxo-2-thioxo-9H-purin-8-yl)cinnamicAcid Nonaethylene Glycol Methyl Ether Ester

(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-6-oxo-2-thioxo-9H-purin-8-yl)cinnamicacid (WO 96/04280, 500 mg, 0.99 mmol) was esterified with nonaethyleneglycol methyl ether by the method of part (b) of Example 24. Titlecompound was isolated as a yellow waxy solid (0.145 g, 20%); ¹H-NMR(DMSO-d,)δ: 8.17 (d, J=8.4, 2, 2 aryl CH), 7.90 (d, J=8.4, 2, 2 arylCH), 7.69 (d, J=15.9, 1, CH=), 6.77 (d, J=16.1, 1, CH=), 4.53 (d, J=7.0,2, CH₂N), 4.40 (d, J=7.0, 2, CH₂N), 4.25 (m, 2, CH₂O), 3.70 (m, 2,CH₂O), 3.6-3.3 (m, 32, 16 CH₂), 3.23 (s, 3, CH₃), 2.4-2.0 (2 m 2, 2CH ofcyclohexyl), 1.80-1.60 and 1.20-1.0 (both m, 20, cyclohexyl CH₂).

Anal. Calcd for C₄₇H₇₃N₄O₁₂S.0.89 H₂O: C, 60.49; H, 7.97; N, 6.00; S,3.44. Found: C, 60.49; H, 7.70; N, 6.31; S, 3.55.

EXAMPLE 27(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamicAcid Nonaethylene Glycol Methyl Ether Ester

To a mixture of(E)-1,3-bis(cyclohexylmethyl)-8-(3-(2-(1H-imidazol-1-ylcarbonyl)vinyl)phenyl)-9H-purin-2,6(1H,3H)-dione(60.75 g, 0.112 mole) and potassium carbonate (31.0 g, 0.225 mole) inacetonitrile (650 mL) was added nonaethylene glycol monomethyl ether,(part (a) of example 10, 57.8 g, 135 mmol). The mixture was refluxed 18h, cooled to ambient temperature, and diluted with chloroform (1200 mL).The chloroform solution was washed with 1N hydrochloric acid (800 mL),water (500 mL), and brine (2×200 mL) and dried (magnesium sulfate.Evaporation of chloroform left crude title compound as a yellow waxysolid. This solid was chromatographed twice on silica gel, first using10% methanol-chloroform and then using 10% methanol-ethyl acetate aseluent, to give the title compound as a yellow waxy solid.Reprecipitation from chloroform-hexanes and drying in vacuo titlecompound as a yellow waxy solid (64.5 g, 65%); ¹H-NMR identical to thesample described in Example 10, part (b).

Anal. Calcd for C₄₇H₇₂N₄O₁₃: C, 62.65; H, 8.05; N, 6.22. Found: C,62.33; H, 7.94; N, 6.25

Pharmaceutical Formulation Examples

In the following Examples, the “active ingredient” may be any compoundof formula (I) or a pharmaceutically acceptable salt or solvate thereofpreferably compound of Examples 2 to 26.

(1) Tablet Formulations

(i) Oral

mg/tablet A B C Active ingredient 25 25 25 Avicel ™ 13 — 7 Lactose 78 47— Starch (maize) — 9 — Starch (pregelatinised, NF15) — — 32 Sodiumstarch glycollate 5 — — Povidone ™ 3 3 — Magnesium stearate 1 1 1 125 8565

(ii) Sublingual

mg/tablet D E Active ingredient 25 25 Avicel ™ — 10 Lactose — 36Mannitol 51 57 Sucrose —  3 Acacia —  3 Povidone ™  3 — Magnesiumstearate  1  1 90 125 

Formulations A to E may be prepared by wet granulation of the first sixingredients with the povidone™, followed by addition of the magnesiumstearate and compression.

(iii) Buccal

mg/tablet Active ingredient 25 Hydroxypropylmethyl 25 cellulose (HPMC)Polycarbophil ™ 39 Magnesium stearate 1 90

The formulation may be prepared by direct compression of the admixedingredients.

(2) Capsule Formulations

(i) Powder

mg/Capsule F G Active ingredient 25 25 Avicel ™ 45 — Lactose 153 —Starch (1500 NF) — 117 Sodium starch glycollate — 6 Magnesium stearate 22 225 150

Formulations F and G may be prepared by admixing the ingredients andfilling two-part hard gelatin capsules with the resulting mixture.

(ii) Liquid fill

mg/Capsule H I Active ingredient  25  25 Macrogol ™ 4000 BP 200 —Lecithin — 100 Arachis oil — 100 225 225

Formulation H may be prepared by melting the Macrogol™ 4000 BP,dispersing the active ingredient in the melt and filling two-part hardgelatin capsules therewith. Formulation I may be prepared by dispersingthe active ingredient in the lecithin and arachis oil and filling soft,elastic gelatin capsules with the dispersion.

(iii) Controlled release

mg/tablet Active ingredient 25 Avicel ™ 123 Lactose 62 Triethylcitrate 3Ethyl cellulose 12 225

The formulation may be prepared by mixing and extruding the first fouringredients and spheronising and drying the extrudate. The dried pelletsare coated with ethyl cellulose as a release controlling membrane andfilled into two-part, hard gelatin capsules.

(3) Intravenous Injection Formulation

(i) % by weight Active ingredients 2% Sodium hydroxide q.s to pH 7 Waterfor Injections to 100%

The active ingredient is taken up in the citrate buffer and sufficienthydrochloric acid added to affect solution and adjust the pH to 7. Theresulting solution is made up to volume and filtered through a microporefilter into sterile glass vials which are sealed and oversealed.

Example G

Powder Capsules for Inhalation

Active Ingredient (0.5-7.0 μm powder) 1.0 mg Lactose (30-90 μm powder)49.0 mg

The powders were mixed until homogeneous and filled into suitably sizedhard gelatin capsules (50 mg per capsule).

Example H

Inhalation Aerosol

Active Ingredient (0.5-7.0 μm powder) 50.0 mg Sorbitan Trioleate 100.00mg Saccharin Sodium (0.5-7.0 μm powder) 5.0 mg Methanol 2.0 mgTrichlorofluoromethane 4.2 g Dichlorodifluoromethane to 10.0 ml

The sorbitan trioleate and methanol were dissolved in thetrichloro-fluoromethane. The saccharin sodium and active ingredient weredispersed in the mixture which was then transferred to a suitableaerosol canister and the dichlorofluoromethane injected through thevalve system. This composition provides 0.5 mg of active ingredient ineach 100 μl dose.

Biological Activity

1) Carrageenan Pleurisy Assay

The antiinflammatory activity of compounds of the invention wasdetermined by the procedure of Vinegar, R, et al., Proc. Soc. Exp. Biol.Med., 1981, 168, 24-32, using male Lewis rats of 150±20 grams. Thecarrageenan dose was 0.075 mg/rat. Pleural exudate was harvested fourhours after injection of carrageenan. Acute antiinflammatory activitywas determined by inhibition of pleural edema and inflammatory cells(neutrophils) from a negative (vehicle) control group.

2) Acetic Acid Colitis Assay

Anti-inflammatory activity of compounds of the invention was determinedin the Acetic Acid Colitis rat model using the procedure of Fretland,D., et al., 1990, 255:572-576 in male Lewis rats 275+25 grams. Compoundswere administered either orally or rectally 24, 16 and 4 hours prior tothe 40 second instillation of 3% acetic acid solution in the proximal 6cm of the colon under light anesthesia. The colon was immediately washedwith 5 cc of saline. 24 hours later the rats were sacrificed and 6 cm ofthe proximal colon was excised weighed for edema. Neutrophilinflammation was determined by measuring MPO levels in the scrapedcolonic mucosa from these rats. Anti-inflammatory activity wasquantitated by inhibition of edema formation and mucosal MPO levelscompared to the negative control group (vehicle).

Results

ACETIC ACID COLITIS (24 hr) CARRAGEENAN ACTIVE DOSE PLEURISY (4 hr)[MG/KG] n LOCAL (ROUTE) (or ED₅₀ (MG/RAT) TISSUE COMPOUND PEG avg.)CELLS EDEMA MPO WEIGHT Dexamethasone 0.02 0.015 0.03 0.17 (ic) (ic)Example 9 CH₃ 10 0.02 0.2 5.0 5.0 (ic) (ic) Example 10 CH₃ 9 0.5 0.5 5050 (po) (po) Example 15 CH₃ 15 or 16 0.4 0.2 50 inactive (po) (50, po)Example 14 CH₃ 41.5 Inactive 0.5 NT NT 0.5 Example 18 OH 13 0.1 0.1 NTNT Example 17 OH 18.9 0.1 0.1 NT NT Example 16 OH 32.2 0.2 0.2 NT NT NT= Not tested ic = Intracolonic administration po = Oral administration

The parent acid i.e. the compound reference example 1 is inactive inboth assays.

3) In Vivo Septic Shock Model: C Parvum/LPS Shock

Male CD-1 mice, 25-30 g (Charles River: Raleigh, N.C.), are injectedintravenously (i.v.) with 100 μg killed C. Parvum (Coparvax; BurroughsWellcome, RTP, N.C.). Ten days later the mice are injected i.v. with 20μg E. Coli 026:B6 lipopolysaccharide (LPS; Difco Labs, Detroit, Mich.)in the presence of the analgesic butorphenol (150 μg per mouse).

The test compounds are dissolved in dimethyl sulphoxide and diluted into0.5% methyl cellulose and then dosed orally 2 hours before the LPS andat the same time as the LPS.

Results

Compound (Oral Dose) Alive/Total % Survivors Control 0/8 0 Example 9 4/850 (75 mg/kg Reference 2/8 25 Example 1 (75 mg/kg)

What is claimed is:
 1. A compound selected from the group consisting of:(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic acid triethylene glycol methyl ether ester;(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic acid polyethylene glycol (n=7.2) methyl ether ester;(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic acid tetraethylene glycol methyl ether ester;(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic acid pentaethylene glycol methyl ether ester;(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic acid hexaethylene glycol methyl ether ester;(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic acid heptaethylene glycol methyl ether ester;(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic acid octaethylene glycol methyl ether ester;(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic acid polyethylene glycol (n=11.7) methyl ether ester;(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic acid hexaethylene glycol ester;(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic acid polyethylene glycol (n=23.9) methyl ether ester;(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic acid polyethylene glycol (n=41.5) methyl ether ester;(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic acid polyethylene glycol (n=15) methyl ether ester;(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic acid polyethylene glycol (n=32.2) ester;(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic acid polyethylene glycol (n=18.9) ester;(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)cinnamic acid polyethylene glycol (n=13) ester;(E)-3-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)benzoic acid nonaethylene glycol methyl ether ester;(E)-2-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-2,6-dioxo-9H-purin-8-yl)benzoic acid nonaethylene glycol methyl ether ester;(E)-4-(1,3-Bis(cyclohexylmethyl)-1,2,3,6-tetrahydro-6-oxo-2-thioxo-9H-purin-8-yl)cinnamic acid nonaethylene glycol methyl ether ester; andpharmaceutically acceptable solvates thereof.